BOOK OF ABSTRACTS 2 CONTENTS INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX INTRODUCTION 3 DEAR COLLEAGUES, as Chair of the Board of Trustees of EUROMAR I want to welcome you all to the EUROMAR meeting 2021. Unfortu- nately, like last year, the pandemic situation does not allow us to meet in person. I am sure you all regret not being at the beautiful Slovenian seaside in Portoroz as much as I do. Within the last year, we all painfully realized how important direct personal interaction is for scientific research, as well as for scientific education. Of course, virtual meetings cannot compensate all aspects of conventional physical meetings. On the other hand, they also offer some new opportunities and possibilities. I am very grateful to Janez Plavec, who agreed in 2019 (well before the THOMAS PRISNER pandemic situation) to organize the EUROMAR meeting 2021 and who courageously and unwaveringly continued — Chair of the Board of Trustees his commitment even in this difficult situation. Organizing an event like the EUROMAR conference under these of EUROMAR circumstances is not only much more work but also a ‘high risk’ endeavor. We all more or less know how ‘normal’ conferences work, but many things are different and have to be different with online meetings. As experimental scientists, thankfully we are all somewhat used to an uncertain situation like this, so we are not afraid to explore the different parameters and are interested in the results. I hope that we will all enjoy this online EUROMAR meeting as much as possible, listen to new and exciting scientific talks, use the opportunity to discuss with poster presenters, keep our interaction with vendors active and enjoy to see each other, if only through the screen of our computers. I would very much like to thank all the sponsors and vendors for their financial support and their solidarity in these special times, the organization and scientific committees for their work and, last but not least, I would like to thank Janez Plavec once again for his vigor and determination to get it going. I wish you all a successful meeting and hope very much to see you in person again soon. INTRODUCTION SPONSORS PROGRAM Best regards, PROGRAM — BY DAY TOPICS Thomas Prisner PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX INTRODUCTION 4 DEAR PARTICIPANTS OF THE EUROMAR 2021 CONFERENCE, DEAR COLLEAGUES, It is a great pleasure to welcome you all on behalf of the organizing changed, including our ability to meet and discuss freely at a con-committee to this year’s EUROMAR conference. A warm welcome ference. Yet, life goes on and the magnetic resonance community to plenary and invited speakers, presenters of promoted talks and has demonstrated, once more, our ability to endure and overcome poster presenters. We are about to start an exciting four-day confer-difficulty. A virtual version of EUROMAR 2021 aims to provide a ence with many contributions from groups all over the world. EU- well-deserved recognition of award winners and especially young JANEZ PLAVEC ROMAR 2021 conference had to be transformed a few times during scientists. The organizers admit that this year’s conference can be — Chair Euromar 2021 past last two years. Local organizers, members of the Slovenian considered an experiment. Every poster presentation is supported NMR centre, are sorry that we were not allowed to gather everyone with a short video presentation in addition to the standard abstract in person and to enjoy your company and private discussions in a that will hopefully stimulate interactions through various media scenic and stimulating as much as the refreshing environment of platforms. You, the participants of the conference, will have the the northern Adriatic. opportunity to provide feedback on these new ways to organize an international event. In early Spring this year a very unfortunate decision had to be made. With all the optimism and enthusiasm needed to organize a major May I personally thank you all for your support and understanding. international conference, we were forced to move EUROMAR 2021 Finally, I want to extend my gratitude to the members of the Interna-to virtual format only. On the positive side, all lectures have been tional Scientific Committee, who have helped to shape the scientific uploaded into the conference platform and are available on de-program, vendors and supporters, who contributed through finan- INTRODUCTION mand. This may be of great advantage for our colleagues in the far cial support, and my colleagues that actively participated in organi- SPONSORS East and far West who will not be forced to stay up very late or to get zation of EUROMAR 2021 conference. Their names are listed on the PROGRAM up very early in order to follow presentation of their favorite speak- conference webpage and in the booklet. A sincere and big apprecia- PROGRAM — BY DAY er. Registered attendees will be able to watch recordings of oral Q tion for their support and contribution. Thank You all. TOPICS & A sessions that will take place on-line at the end of each session. PRIZE LECTURES Vendor webinars will be accessible for later viewing as well. New PLENARY LECTURES communication methods now allow everyone to get in touch with Looking forward to meeting you all in the near future live, TUTORIAL LECTURE anyone with a click and a short note. INVITED AND PROMOTED LECTURES Janez Plavec POSTERS Our lives and everyday activities were affected in so many ways. ADVERTISERS Interactions, teaching, sharing excitement over new results, etc. AUTHOR INDEX have been taking new forms as the very essence of our society has INTRODUCTION 5 ORGANIZING INTERNATIONAL SCIENTIFIC LOCAL ORGANIZING BOARD OF TRUSTEES: PROGRAM COMMITTEE: COMMITTEES COMMITTEE: • Thomas Prisner • Janez Plavec, • Janez Plavec • Arno Kentgens National Institute of Chemistry, Slovenia • Martina Lenarčič-Živković • Sabine Hediger • Thomas Prisner, • Maja Marušič • Wiktor Kozminski University of Frankfurt, Germany • Petra Jaksetič • Matthias Ernst • Marina Bennati, • Anamarija Anja Novak • Tatyana Polenova Max Planck Institute and • Marko Trajkovski • Luisa Ciobanu University of Göttingen, Germany • Primož Šket • Oscar Millet • Patrick Giraudeau, • Gregor Mali • Patrick Giraudeau University of Nantes, France • Damjan Makuc • Sharon Ashbrook • Luisa Ciobanu, • Peter Podbevšek • Dimitrios Sakellariou CEA-Saclay, France • Maria Toplishek • Gunnar Jeschke • Issabela Felli, • Anita Kotar • Janez Plavec University of Florence, Italy • Vojč Kocman • Andrew Webb • Anne Lesage, • Matjaž Polak • Marc Baldus Institute of Analytical Sciences, • Janez Dolinšek • Roberta Pierattelli Lyon, France • Tomaž Apih • Daniel Topgaard • Janez Dolinšek, • Denis Arčon • Melanie Britton INTRODUCTION Institute Jožef Stefan, Slovenia • Martin Klanjšek SPONSORS • Wiktor Kozminski, • Aleš Mohorčič PROGRAM University of Warsaw, Poland • Igor Serša PROGRAM — BY DAY • Lukas Trantirek, • Matej Preglej TOPICS Masaryk University, Czech Republic • Boštjan Zalar PRIZE LECTURES • Georgios Papavassiliou, • Andrej Zorko PLENARY LECTURES National Centre of Scientific Research TUTORIAL LECTURE "Demokritos", Greece INVITED AND PROMOTED LECTURES • Tomaž Apih, POSTERS Institute Jožef Stefan, Slovenia ADVERTISERS • Yishay Manassen, AUTHOR INDEX Ben Gurion University of the Negev, Israel SPONSORS 6 SPONSORS INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX SPONSORS 7 SPONSORS INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PROGRAM 8 MONDAY, JULY 5 TUESDAY, JULY 6 WEDNESDAY, JULY 7 THURSDAY, JULY 8 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PROGRAM 9 MONDAY, JULY 5 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PROGRAM 10 TUESDAY, JULY 6 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PROGRAM 11 WEDNESDAY, JULY 7 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PROGRAM 12 THURSDAY, JULY 8 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX TOPICS 13 PROGRAM • BIOMOLECULES AND INTERACTIONS TOPICS • BIOSOLIDS • BIOMOLECULAR DYNAMICS • INTERGRATED STRUCTURAL BIOLOGY: NMR IN HAND WITH COMPLEMENTARY METHODS • IN-CELL MAGNETIC RESONANCE • SMALL MOLECULES • DRUG DESIGN AND COMBAT AGAINST COVID-19 • METABOLOMICS • HYPERPOLARIZATION • FIELD-CYCLING NMR RELAXOMETRY • MRI IN MATERIAL SCIENCE AND BIOMEDICAL APPLICATIONS • ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS • ORGANIC AND COMPOSITE SOLIDS INTRODUCTION SPONSORS • METHODS DEVELOPMENT PROGRAM • BENCHTOP AND LOW-FIELD PROGRAM — BY DAY TOPICS • EPR IN BIOMOLECULAR AND MATERIAL SCIENCE PRIZE LECTURES • FRONTIERS IN MAGNETIC RESONANCE PLENARY LECTURES TUTORIAL LECTURE • NMR OF QUANTUM MATERIALS INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX 14 PRIZE LECTURES PR001 PRIZE LECTURE / AWARD FOR AMPERE PRIZE 15 ANTOINE LOQUET STRUCTURAL BIOLOGY OF PROTEIN ASSEMBLIES CNRS, IECB, CBMN, AND PATHOGEN CELL SURFACE BY SOLID-STATE NMR University of Bordeaux, Bordeaux, France SPECTROSCOPY We will present the NMR activity carried out in the laboratory, focusing on recent and unpublished results: • solid-state NMR-based structure determination of amyloid fibrils using ¹H detection and ¹H-¹H distance re- straints at fast magic-angle spinning; • multidimensional ¹H-detected solid-state NMR of protein assemblies at fast magic-angle spinning combined with DNP; • development of 1H detection at fast magic-angle spinning to study the surface of pathogens. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PR002 PRIZE LECTURE / AWARD FOR ANDREW PRIZE 16 T. REID ALDERSON¹,² PROTEIN FOLDING INVESTIGATED BY NMR SPECTROSCOPY 1Department of Chemistry, Physical and Theoretical In my thesis research, I used NMR spectroscopy, which simultaneously provides atomic-level insight into both the structures and dynam-Chemistry Laboratory, University ics of biomolecules, to investigate two fundamental aspects of biochemistry: how do proteins fold from linear chains of amino acids into of Oxford, South Parks Road, complex three-dimensional structures, and how do the class of proteins known as molecular chaperones prevent pathological protein mis-Oxford, UK 2Laboratory of Chemical Physics, folding and aggregation? Dysregulated protein folding contributes to the etiology of protein misfolding diseases, including Alzheimer’s, National Institutes of Health, Parkinson’s and type II diabetes. Understanding the precise molecular mechanisms that underpin these processes could guide the design Bethesda, MD, USA of future therapeutics that help mitigate protein misfolding diseases. During my PhD, I determined a redox-controlled mechanism that regulates the function of the molecular chaperone HSP27 [1, 2], and I elucidated the structural basis for mutations in HSP27 that cause incurable motor neuropathies [3]. I also contributed to the development of novel NMR methodology involving rapid pressure changes to study protein folding at atomic resolution on the millisecond timescale, enabling unprecedented insight into the early stages of folding and misfolding [4-6]. By performing complementary static pressure experiments, I quantified the compaction of an unfolded polypeptide chain as the difference in free energy between the unfolded and folded states is lowered [7]. In natively unfolded and denatured proteins, I studied the formation of cis-proline bonds, which often must isomerize to the trans conformation before folding can occur, and I found that model peptides overestimate the fraction of the cis conformation [8]. Finally, I aided in the development of NMR software that automatically assigns methyl resonances from highly deuterated, selectively methyl-13CH labeled proteins up to 1 MDa in aggregate mass, with assignments obtained based on a comparison of through-space experimental 3 restraints (NOEs) and high-resolution structures [9, 10]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS References: [1] Alderson TR, Roche J, Gastall HY, Dias DM, Pritišanac I, Ying J, Bax A, Benesch JLP, Baldwin AJ, Nat. Commun. , 2019, 10, 1068. [2] Alderson TR, Ying J, Bax A, Benesch JLP, Baldwin PRIZE LECTURES AJ, J. Mol. Biol. , 2020, 432, 3033-3049. [3] Alderson TR, Adriaenssens E, Asselbergh B, Pritišanac I, Van Lent J, Gastall HY, Walti MA, Louis JM, Timmerman V, Baldwin AJ, Benesch JLP. EMBO PLENARY LECTURES J. , 2021, 40, e103811. [4] Alderson TR, Charlier C, Torchia DA, Anfinrud P, Bax A, J. Am. Chem. Soc. , 2017, 139, 11036-11039. [5] Charlier C, Alderson TR, Courtney JM, Ying J, Anfinrud P, Bax A, TUTORIAL LECTURE PNAS, 2018, 115, E4169-E4178. [6] Charlier C, Courtney JM, Alderson TR, Anfinrud P, Bax A, J. Am. Chem. Soc. , 2018, 140, 8096-8099. [7] Ramanujan V, Alderson TR, Pritišanac I, Ying J, Bax A, J. Mag. Reson. , 2020, 312, 106701. [8] Alderson TR, Lee JH, Charlier C, Ying J, Bax A, ChemBioChem, 2018, 19, 37-42. [9] Pritišanac I, Degiacomi MT, Alderson TR, Carneiro MG, Ab E, Siegal G, INVITED AND PROMOTED LECTURES Baldwin AJ, J. Am. Chem. Soc. , 2017, 139, 9523-9533. [10] Pritišanac I, Wurz JM, Alderson TR, Guntert P, Nat. Commun. , 2019, 10, 4922. POSTERS Acknowledgements: I am grateful to my supervisors, Dr. Ad Bax, Prof. Justin L. P. Benesch, and Prof. Andrew J. Baldwin for their support, advice, and inspiration during my thesis research. ADVERTISERS I thank Prof. Dame Carol V. Robinson and Dr. G. Marius Clore for sharing their laboratory facilities and reagents. I am indebted to Prof. Christina Redfield and Prof. D. Flemming Hansen for AUTHOR INDEX their careful examination of my thesis and for providing helpful corrections. PR003 PRIZE LECTURE / AWARD FOR ERNST PRIZE 17 JAN HENRIK DISSOLUTION DYNAMIC NUCLEAR POLARIZATION ARDENKJÆR- LARSEN Hyperpolarized Metabolic MR is a novel medical imaging modality that offers exceptional possibilities to follow changes in metabolism in vivo in real time [1]. The method is enabled by a more than 10,000 fold enhancement [2] Technical University of Denmark, Department of Health of the signal from metabolic contrast agents that probe central metabolic pathways. The contrast agent is typically Technology, Kongens Lyngby, enriched in 13C and polarized by dissolution Dynamic Nuclear Polarization (dDNP). The contrast agent circulates via Denmark the vasculature to the tissue of interest, where it is taken up by the tissue cells and metabolized into specific products. MR is unique in several ways: 1) it already provides anatomical and morphological images with high resolution and contrast based on the tissue water protons, 2) it is does not expose the patient to any ionizing radiation, and 3) it is a spectroscopic method that allows quantification of the individual metabolites. The first tracer in clinical development is 13C-pyruvate. Pyruvate is at a pivotal point in glycolysis and allows us to directly probe the Warburg effect in cancer through the elevated lactate-to-pyruvate ratio, but also other pathologies. The hope is that more accurate diagnosis and staging can be made, and that the method will provide an early read-out of response to treatment. The first clinical studies have been published [3–6] with encouraging results. In this talk I will review dDNP of 13C and 1H with trityl and UV radicals with the aim of reaching higher polarization in a faster manner. Direct polarization of 13C with trityl is robust and efficient, however, subtle differences in the chemical structure of the trityl and change of matrix, affects the DNP efficiency. In contrast, UV radicals are labile, and quench in the solid state well below room temperature, which may provide a means to hyperpolarized solids with long relaxation for storage and transportation. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] J. Kurhanewicz, D.B. Vigneron, J.H. Ardenkjaer-Larsen, et al., Neoplasia 2019, 21, 1–16; [2] J.H. Ardenkjær-Larsen, B. Fridlund, A. Gram, et al., INVITED AND PROMOTED LECTURES Proc. Natl. Acad. Sci. U. S. A. 2003, 100, 10158–63; [3] S.J. Nelson, J. Kurhanewicz, D.B. Vigneron, et al. , Sci. Transl. Med. 2013, 5, 198ra108; [4] C.H. Cunningham, POSTERS J.Y. Lau, A.P. Chen, et al, Circ. Res. 2016, 116, 309769; [5] R. Aggarwal, D.B. Vigneron, J. Kurhanewicz, Eur. Urol. 2017, 72, 1028–1029; [6] V.Z. Miloushev, K.L. ADVERTISERS Granlund, R. Boltyanskiy, et al. , Cancer Res. 2018. AUTHOR INDEX Acknowledgements: Danish National Research Foundation (DNRF124). PR004 PRIZE LECTURE / AWARD FOR ERNST PRIZE 18 LUCIO FRYDMAN BASKING IN ERNST’S LEGACY: Department of Chemical and PULSES, SPECTRA, IMAGES Biological Physics, Weizmann Institute of Science, Rehovot, Israel; lucio.frydman@ Even for those of us that never had the privilege to work with Richard Ernst (1937 – 2021), the man and his perso-weizmann.ac.il na stand out among the greatest of our generation. In fact Ernst’s greatness can probably be best appreciated by none better than us, the practitioners of Nuclear Magnetic Resonance. His experiments, his formalisms and deriva-tions: they all carry a stamp of ingenuity, depth and rigorousness that is typically Ernst’s. Whether revisiting a previous proposition or making a completely new one, whether being the sole senior author or pushing the envelope of Science in a collaboration, most of us would recognize “an Ernst” just by reading its pages. In the present short talk I will present how three such classical “Ernsts” –the Ernst/Anderson pulsed approach to high resolution NMR, his proposition to collect 3D images non-invasively by Fourier NMR Zeugmatography, and the Jeener/Ernst proposition to unravel complex information by multidimensional NMR– have molded our own research. I will specifically focus on how pulsed NMR led us to propose an alternative wat to collect 1D NMR spectra by spatiotemporal encoding, opening in turn the possibility to acquire arbitrary multidimensional NMR spectra / images in a single scan. The talk will center mostly on the latter’s potential, highlighting ongoing preclinical and clinical projects dealing with understanding and diagnosing breast and pancreatic cancers, following fetal and neonatal development in normal and abnormal pregnancies, and tackling new functional / structural aspects of the brain with very high definition. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX 19 PLENARY LECTURES PL001 PLENARY LECTURE / BIOMOLECULES AND INTERACTIONS 20 ROBERTA PIERATTELLI1,2 UN-STRUCTURAL BIOLOGY BY NMR SPECTROSCOPY 1Magnetic Resonance Center (CERM), University of Florence, Italy Intrinsically disordered proteins (IDPs) and regions (IDRs) that do not adopt a well-defined three-dimensional struc-2Department of Chemistry “Ugo Schiff”, ture under physiological conditions are now well recognized in structural biology. The last two decades have seen University of Florence, Italy increasing evidence for the involvement of IDPs and IDRs in many complex biological pathways, complementing globular proteins’ molecular functions. Historically understudied, intrinsic protein disorder is nowadays central in an increasingly large number of studies. The structural properties of highly dynamic polypeptides cannot be captured in ordered crystals, preventing them to be suitable targets for crystallographic studies. Thus, nuclear magnetic resonance (NMR) spectroscopy plays a crucial role in the characterization of highly flexible regions in multi-domain proteins and entire proteins characterized by the lack of a 3D structure. The high flexibility has several consequences on the NMR spectroscopic parameters that, if properly handled, can give precious information. I will illustrate how NMR can help in describing the importance of intrinsic disorder to encode in a relatively short polypeptide many functional modules and to orchestrate protein-protein interactions. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PL002 PLENARY LECTURE / NMR OF QUANTUM MATERIALS 21 PHILLIPE MENDELS NMR IN QUANTUM MATERIALS: Université Paris-Saclay , CNRS, FROM MILESTONES TO THE INTRIGUING “KAGOME” CASE Laboratoire de Physique des Solides, Orsay, France In recent years, there has been a growing interest in material sys- search, still very active, for both new theories and materials with a tems where quantum effects play a major role in emergent func- kagome geometry. In my talk, I will mainly focus on a few among a tionalities or as a seed for novel concepts. This vast field of research growing number of kagome compounds where such a state is now from fundamental to applications encompasses superconductors, stabilized but resists to a definite interpretation. Herbertsmithite, graphene, topological insulators, Weyl semimetals, quantum spin ZnCu (OH) Cl , which has been known since 2005 as one of the best 3 6 2 liquids… representative of that spin liquid physics has been at the center of our activity on quantum materials in Orsay, Recently, working on My talk will be on the fundamental side, focusing on systems where high quality single crystals considerably improved the accuracy of quantum effects are manifest in spin systems. Very simple Hamil- NMR measurements and its ability to address fundamental issues tonians accounting for the interactions between spins are at work, such as the class of models relevant to the description of the ground yet many uncovered phases can be generated by a combination of a state. Despite the existence of out-of-kagome planes defects which reduced dimensionality, of the geometry of the lattice and of frus-mask the intrinsic physics, one can indeed isolate the 17O NMR spec- tration. The topic has expanded a lot over the last 30 years. tral signature of kagome spins S=1/2 at Cu2+ sites. This illustrates After introducing the field, I’ll focus on the case of quantum spin all the power of NMR as a local probe aimed at imaging the cen-liquids. This quantum disordered ground state was proposed by An- tral physics at work through a wise selection of the probe nucleus. derson in 1973 as an alternative to the standard antiferromagnetic The shift measurements give access to the local susceptibility and INTRODUCTION state “à la Néel”. The discovery of high temperature superconduc- spins dynamics, through the prism of T relaxation, shed light on 1 SPONSORS tivity in 1986 acted as a revival for the search of such a quantum excitations in a low energy range specific to NMR. This a common PROGRAM spin liquid, based on his ideas again, of a resonating valence bond working scheme to all NMR groups studying quantum materials. I’ll PROGRAM — BY DAY state. Soon after, the antiferromagnetic kagome lattice decorated finally complement my presentation by showing how ESR can help TOPICS with quantum spins was proposed as the best candidate for stabiliz- in characterizing deviations to the Heisenberg model which have to PRIZE LECTURES ing such a state in dimension higher than one which triggered the be taken into account in the comparison to models. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] B. Keimer,J. Moore, Nature Physics 2017, 13, 1045. [2] L. Savary, L. Balents, Rep. Prog. Phys.. 2017, 80, 016502. [3] P. Mendels, F. Bert, C.R. Phys 2016, 17, 455 [4] M. Fu, T. Imai, T.-H. Han , Y.S. Lee et al., Science. ADVERTISERS 2015, 350, 655. [5] P. Kuntia et al., Nature Physics 2020, 16, 469. [5] A. Zorko et al. , Phys.Rev. Lett. 2008, 101, 026405. AUTHOR INDEX Acknowledgements: French Agence Nationale de la Recherche, Grant No ANR-18-CE30-0022. PL003 PLENARY LECTURE / IN-CELL MAGNETIC RESONANCE 22 GARY J. PIELAK PROTEIN & PROTEIN-COMPLEX STABILITY Department of Chemistry, IN LIVING CELLS University of North Carolina at Chapel Hill, Chapel Hill, NC, USA The crowded and complex environment in cells is predicted to affect protein behavior compared to dilute buffer. My laboratory and our collaborators are examining crowding effects on the stability of proteins and their complexes in cells and under physiologically relevant crowded conditions using NMR. A challenge in these endeavors is detecting the test protein in a sea of crowders. NMR is ideally suited to overcome this challenge and provide high quality data on folded- and unfolded- proteins as well as free and bound forms of complexes. I will focus on equilibrium data acquired in living Escherichia coli cells, Xenopus laevis and Danio rerio oocytes as well as in vitro in concentrated cosolute solutions using 19F NMR. The cosolutes include synthetic polymers and their monomers, other proteins and lyophilized cytosol. The results show that crowding affects folding and binding in ways not always correctly predicted by simple theory. The differences point to opportunities for theoretical efforts and simulations. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: A. J. Guseman & G. J. Pielak, Chapter 12 protein stability and weak intracellular interactions. In-cell NMR spectroscopy: From molecular sciences to cell biology, eds. Y. Ito, V. Dötsch, and M. Shirakawa (The Royal Society of Chemistry), 2020, 188-206; S. S. Stadmiller, J. S. Aguilar, C. Waudby, and G. J. INVITED AND PROMOTED LECTURES Pielak, Biophys. J. 2020, 118, 2337-2348; S. L. Speer, W. Zheng, X. Jiang, I.-T. Chu, A. Guseman, M. Liu, G. Pielak, and C. Li, Proc. Natl. Acad. Sci. USA 2021, 118, POSTERS e2019918118; J. Thole, T. Fadero, J. Bonin, S. Stadmiller, J. Giudice, and G. Pielak, Biochemistry 2021, 60:451-459. ADVERTISERS Acknowledgements: I thank the US National Science Foundation (MCB 1410854) and the U.S.-Israel Binational Science Foundation (BSF 2017063) for AUTHOR INDEX supporting my group’s efforts on crowding. PL004 PLENARY LECTURE / SMALL MOLECULES 23 WILLIAM GERRARD, NMR-BASED STRUCTURE ELUCIDATION – SIYING ZHONG, DENSITY FUNCTIONAL THEORY, MACHINE LEARNING CRAIG BUTTS School of Chemistry, AND A LOT OF DATA University of Bristol, Bristol, United Kingdom Interpreting NMR spectra has become somewhat of an art form in chemistry, particularly when applied to molecu- lar structure elucidation. Skilled practitioners might pore over several, often complex, NMR spectra for hours, days and even weeks to work out the connectivity (2D structure), stereochemistry and conformation (3D structure) of challenging new molecules. Our team develop tools that help with the steps in this process: creating new NMR experiments that provide dif- ferent or more quantitative information than existing methods; applying quantum chemical calculations (usually Density Functional Theory) to accurately predict the NMR properties of candidate 3D molecular structures; developing machine learning tools that can accelerate predictions with 3D structural relevance by 10,000-fold [1] and thus potentially allow us to screen hundreds of thousands of molecular structures to find good fits to the experimental NMR spectra. The potential (and limits!) of these approaches will be illustrated by the case of the structure reassignment of the baulamycin natural products [2] - molecules with 128 diastereoisomers, wach with around 1 billion potential 3D conformations - which we solved by combining elegant chemical synthesis (which I won’t talk about much) with quantitative NMR and computational methods (which I will). INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] W. Gerrard, L. A. Bratholm, M. J. Packer, A. J. Mulholland, D. R. Glowacki, C. P. Butts, Chem. Sci. 2020, 11, 508-515. [2] J. Wu, AUTHOR INDEX P. Lorenzo, S. Zhong, M. Ali, C. P. Butts, E. L. Myers, V. K. Aggarwal, Nature. 2017, 547, 436-440. PL005 PLENARY LECTURE / FIELD-CYCLING NMR RELAXOMETRY 24 FABIEN FERRAGE MULTISCALE DYNAMICS FROM HIGH-RESOLUTION Laboratoire des Biomolécules, RELAXOMETRY AND TWO-FIELD NMR LBM, Département de chimie, École normale supérieure, PSL University, Sorbonne Université, High magnetic fields have empowered NMR spectroscopists to probe large complex systems with atomic resolu- CNRS, Paris, France tion. Yet, investigating dynamics at very high magnetic fields is often challenging. Line broadening from chemical exchange worsen dramatically while relaxation becomes uninformative about nanosecond motions at very high fields. We have developped, in collaboration with Bruker Biospin, a series of instruments that couple the sensitivity and resolution of high magnetic fields with the evolution of nuclear spins at a lower fields, down to a few mT. Here, we will show how coupling high- and low-field NMR has allowed us to decipher nanosecond motions in protein side chains [1-2], improve models of motions, detect transient interactions in complex mixtures [3] and observe spin systems under intermediate chemical exchange, with spectra broadened beyond detection at high magnetic fields [4]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS References: [1] S. F. Cousin, P. Kadeřávek, N. Bolik-Coulon, Y. Gu, C. Charlier, L. Carlier, L. Bruschweiler-Li, T. Marquardsen, J.-M. Tyburn, R. Brüschweiler and PRIZE LECTURES F. Ferrage, J. Am. Chem. Soc. 2018, 140, 13456–13465. [2] P. Kadeřávek, N. Bolik-Coulon, S. F. Cousin, T. Marquardsen, J.-M. Tyburn, J.-N. Dumez, F. Ferrage, J. PLENARY LECTURES Phys. Chem. Lett. 2019, 10, 5917-5922. [3] Z. Wang, V. Ghini, P. Kadeřávek, S. Pisano, M. Zachrdla, P. Pelupessy, M. Kazmierczak, T. Marquardsen, J.-M. Tyburn, TUTORIAL LECTURE G. Bouvignies, G. Parigi, C. Luchinat, F. Ferrage, ChemRxiv. Preprint. 2021 https://doi.org/10.26434/chemrxiv.13603301.v2. [4] S. F. Cousin, P. Kadeřávek, B. Haddou, C. Charlier, T. Marquardsen, J.-M. Tyburn, P.-A. Bovier, F. Engelke, W. Maas, G. Bodenhausen, P. Pelupessy, F. Ferrage, Angew. Chem. Intl. Ed. 2016, 55, INVITED AND PROMOTED LECTURES 9886-9889 POSTERS Acknowledgements: This research has received funding from the European Union’s Horizon 2020 Program, European Research Council (ERC) Grant ADVERTISERS agreement 825404 (RELAX-MAX), and FET-Open Grant agreement 899683 (HIRES-MULTIDYN) as well as STSM Grants from the EURELAX COST Action AUTHOR INDEX CA15209. PL006 PLENARY LECTURE / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 25 HELEN GRÜNINGER, SOLID-STATE NMR STUDIES OF MATERIALS WOUTER FRANSSEN, FOR ENERGY CONVERSION AND STORAGE HONGTAO QU, SANDER LAMBREGTS, Research and development in energy conversion and storage from been widely used in oxide electrolytes to stabilize the structure and SHRESTHA BANERJEE, renewable sources is increasingly important. As the function of enhance the Li ionic conductivity. We investigate Al-incorporation ERNST VAN ECK, these materials relates to their structure and dynamics, NMR has a in β-Li PS interpretation of the NMR spectra provides insights in 3 4, GILLES DE WIJS, relevant role to play in this research. the effect of Al-doping on the structure and dynamics in Li-Al-P-S ARNO P.M. KENTGENS systems. Alternatively, nanoconfined lithium borohydride shows In the last decade, interest in hybrid halide perovskite materials has high lithium-ion mobility at ambient temperatures. LiBH in silica Radboud University, Institute for skyrocketed due to their remarkable photoluminescent and photo- 4 Molecules and Materials, Nijmegen, nanopores is clearly divided into two distinct fractions at all tem- voltaic (PV) properties. Here NMR is particularly attractive as almost The Netherlands peratures, one being highly dynamic. NMR reveals this is not just all isotopes that constitute the materials are accessible [1]. Compo- due to the nano-confinement, but there is a specific interaction sitional variation of the (organic) cations (A) or halides (X) mixed with the silica pore walls. in A+B +X results in desired effects in optimizing their optoelectric 2 3 properties and improving their stability. We study cation dynamics, Finally, we study rare earth oxyhydrides, such as YO H that x (3-2x), compositional variations and local ordering using solid-state NMR, change color and transparency when irradiated with UV light. Al-NQR, XRD and DFT calculations. Mixing halides influences cation though their optical properties have been studied extensively, the dynamics. The halides also display intriguing dynamics themselves understanding of the relationship between photochromism, chem- allowing us to monitor spontaneous mixing of MAPbI and MAPb- ical composition, structure, and in particular the role of hydrogen, is 3 Br parent compositions. limited [3]. As these materials are grown as 1 3 mm thin films they are INTRODUCTION difficult to characterize by NMR. Nevertheless, we could quantify the All-solid-state lithium batteries are recognized as promising next SPONSORS hydrogen content, get insight in hydride dynamics and identify dif- generation energy storage techniques as they have the potential to PROGRAM ferent yttrium environments in the lattice. A better understanding provide higher energy density, assured safety, prolonged lifespan PROGRAM — BY DAY of the structure function relation can lead to applications in smart and easy processability. To realize them, high ion-conductive solid TOPICS windows or a novel class of electrolytes based on H– ion conductivity. electrolytes have to be developed. NMR is deployed to study Li+ dy- PRIZE LECTURES namics and characterize local structure [2]. Aluminum doping has PLENARY LECTURES TUTORIAL LECTURE References: [1] a) W.M.J. Franssen, A.P.M. Kentgens, Solid State Nucl. Magn. Reson. 2019, 100, 36– 44 b) L. Piveteau, V. Morad, and M.V. Kovalenko, J. Am. Chem. Soc. 2020, 142, 19413–19437 INVITED AND PROMOTED LECTURES [2] O. Pecher, J. Carretero-González, K.J. Griffith, C.P. Grey, Chem. Mater. 2017, 29, 213–242. [3] S. Cornelius, G. Colombi, F. Nafezarefi, H. Schreuders, R. Heller, F. Munnik, and B. Dam, J. Phys. Chem. Lett. 2019, 10, 1342–1348. POSTERS Acknowledgements: We thank our collaboration partners at the University of Bayreuth, University of Twente, Qingdao Energy Storage Research Institute, Chinese Academy of Sciences, ADVERTISERS Utrecht University, Technical University Delft and Radboud University Nijmegen. Support from the Dutch Research Council (NWO), Deutsche Forschungsgemeinschaft (DFG), China AUTHOR INDEX Scholarship Council (CSC) is gratefully acknowledged. PL007 PLENARY LECTURE / HYPERPOLARIZATION 26 LUCIO FRYDMAN SENSITIVITY ENHANCED NMR SPECTROSCOPY Department of Chemical and WITHOUT DNP? C’EST POSSIBLE! Biological Physics, Weizmann Institute, Rehovot, Israel Chemical exchange saturation transfer, or CEST, is a central tool in modern metabolic imaging. CEST exploits the fast exchanges that labile hydrogens undergo with water in order to amplify the signatures of hard-to-observe metabolites. At the same time, it has often been assumed that fast exchanges with the solvent complicate or altogether preclude the observation of the labile sites. In the present study we discuss how exchanges with the solvent can actually be put to good use in order to enhance certain valuable molecular signatures –particularly those involving NOE and TOCSY correlations between labile and non-labile protons in biomolecules. Using relatively simple manipulations tunable by straightforward calibrations, it is shown that solvent exchanges can be used to amplify imino, amino, amide and hydroxy peaks in the 2D NOESY/TOCSY NMR spectra of nucleic acids, proteins and saccharides, by factors ranging from 2 to 10-fold. Even larger gains in sensitivity per unit time –up to two orders of magnitude– can be attained when the spectra are sparse, in which case the information can be efficiently encoded via selective manipulations. Heteronuclear information can also be incorporated into this kind of experiments, leading to high resolution and enhanced sensitivity, in minimal acquisition times. In fact solvent exchanges are not the sole mechanism capable of amplifying NMR spectra in such manner: also spin-diffusion effects among abundant spins, can be used to a similar end when trying to sensitize the spectra of dilute, insensitive species. The talk will conclude with a demonstration of how this can be brought to bear in order to impart high sensitivity/unit_time gains in wideline solid state NMR of 15N, 17O, 33S and other “tough” nuclides. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PL008 PLENARY LECTURE / MRI IN MATERIAL SCIENCE AND BIOMEDICAL APPLICATIONS 27 NOAM SHEMESH INSIGHTS INTO MICRO-ARCHITECTURE FROM DIFFUSION- Champalimaud Research, DRIVEN CORRELATION TENSOR MRI Champalimaud Centre for the Unknown, Lisbon, Portugal Diffusion MRI (dMRI) plays an important role in characterising mi- In the talk, we will explore a new general approach – termed “corre- cro-architectural features in diverse systems ranging from porous lation tensor MRI” (CTI) [10] – for resolving the different diffusional media [1,2] to in-vivo tissues [3]. Non-gaussian diffusion effects, typ-kurtosis sources. CTI [10] is based on displacement correlation ten- ically quantified from diffusional kurtosis [4], have been shown to sor theory [12] and double-diffusion encoding (DDE) [13] waveforms, provide higher sensitivity towards microstructure than their diffu-which enable the explicit measurement of the Z displacement cor- sion tensor counterparts [5]; however, the specificity of diffusional relation tensor [12] as well as the conventional W kurtosis tensor [5]. kurtosis is limited [6] since diffusion non-gaussianity can arise from Hence, the different kurtosis sources can be disentangled without fundamentally different sources. In particular, diffusional kurtosis relying on strong model (or otherwise unverifiable) assumptions can arise from [7,8]: anisotropy of diffusion tensors (K ); variance (n.b. exchange remains a potential confounding factor,). CTI the- aniso in the magnitude of diffusion tensors (K ); and restriction effects [9] ory and validation will be presented, followed by the first imaging iso ( e.g. , full or partial restriction, tortuosity, surface-to-volume effects, results in ex-vivo and in-vivo rat brains revealing striking K , K aniso iso diffusion in heterogeneous cross-sections, etc.), here collevtively and µK contrasts. The first application of CTI for characterizing is-termed microscopic kurtosis (µK). Exchange can also modulate the chemia reveals insights into potential underlying biological corre-extent of the different sources. Contemporary methods based on lates, including neurite beading and edema. Higher CTI sensitivity conventional single-diffusion-encoding (SDE) NMR or MRI cannot towards the ischemic lesion and potential for resolving its hetero-resolve these underlying kurtosis sources, as they are all conflated geneity were observed. All these features bode well for future da- INTRODUCTION in the MR diffusion-driven signal decay [10]. Valiant attempts have vances in CTI methodology and for applications in diverse porous SPONSORS been made for resolving isotropic and anisotropic kurtosis sourc- systems. PROGRAM es [7,8]; however, strict assumptions are invoked, namely, diffusion PROGRAM — BY DAY time-indepdenence [11] and identically zero µK [10] – assumptions TOPICS which likely are not met in many realistic systems. PRIZE LECTURES PLENARY LECTURES References: [1] Y.Q. Song et al., J. Chem. Phys.. 2009, 128, 052212. [2] P. Callaghan, et al. Nature. 1991, 351, 467-469. [3] S. Mori, J. Zhang, Neuron. 2006, 51, 527-539. [4] J.H. Jensen et al. Magn. TUTORIAL LECTURE Reson. Med. 2005, 53, 1432-1440. [5] F. Grinberg et al. NMR Biomed. 2012, 25, 1295-1304. [6] S.U. Rudrapatna et al. Neuroimage. 2014, 97, 363-373. [7] J.P. de Almeida Martins, D. Topgaard. Phys. Rev. Lett. 2016, 116, 087601. [8] F. Szczepankiewicz, et al. Neuroimage. 2015, 104, 241-252. [9] L.N. Burcaw et al. Neuroimage. 2015, 114, 18-37. [10] R.N. Henriques et al. Neuroimage. 2020, 221, INVITED AND PROMOTED LECTURES 116605. [11] S.N. Jespersen et al., J. Magn. Reson 2019, 300, 84-94. [12] S.N. Jespersen NMR Biomed. 2012, 813-818. [13] Y. Cheng, D.G. Cory, J. Am. Chem. Soc. 1999, 121, 7935-7936. POSTERS Acknowledgements: European Research Council (ERC-StG 679058); Champalimaud Centre for the Unknown vivarium (a CONGENTO facility financed by Lisboa Regional Operational ADVERTISERS Programme (Lisboa 2020), project LISBOA01-0145-FEDER-022170); Champalimaud Platforms. The author is grateful to Prof. Dr. Sune N Jespersen, Dr. Rafael Neto Henriques, Dr. Andrada AUTHOR INDEX Ianus, Ms. Rita Alves, Mr. Jonas Olesen, Ms. Beatriz Cardoso, Dr. Cristina Chavarrias, and Dr. Leevi Kerkelä for their profound contributions to CTI. PL009 PLENARY LECTURE / INTEGRATED STRUCTURAL BIOLOGY 28 NATALIYA DANILENKO1, HIGH MOLECULAR-WEIGHT COMPLEXES IN THE LUKAS LERCHER1, REGULATION OF GENE EXPRESSION: A VIEW BY FRANK GABEL3, JOHN KIRKPATRICK1,2, INTEGRATIVE STRUCTURAL BIOLOGY TERESA CARLOMAGNO1,2 1 The Regulator of Ty1 Transposition protein 106 (Rtt109) is a fungal histone acetyltransferase required for histone BMWZ and Institute of Organic Chemistry, Leibniz University of H3 K9, K27 and K56 acetylation. These acetylation sites have been linked to processing and folding of nascent H3 Hannover, Hannover, Germany and play an integral role in replication- and repair-coupled nucleosome assembly. Rtt109 is unique in its activation, 2NMR-based Structural Biology, performed by two structurally unrelated histone chaperones, Asf1 and Vps75. These proteins stimulate Rtt109 activ-Helmholtz Centre for Infection Research, Braunschweig, Germany ity via different mechanisms [1]. Rtt109–Asf1 association has been proposed to be responsible for K56 acetylation, 3Université Grenoble Alpes, Institut de while the Rtt109–Vps75 interaction is required for K9 acetylation [2,3]. Biologie Structurale, Grenoble, France In our work we find that Rtt109, Vps75 and Asf1 are capable of assembling as a previously uncharacterized complex onto the substrate H3-H4 dimer. Using an integrative structural biology approach based on a powerful combination of solution state NMR and small angle neutron scattering (SANS) we solve the structure of this complex and provide a mechanistic explanation for the enzyme activity [4]. We show that Vps75 promotes acetylation of residues in the H3 N-terminal tail by engaging it in fuzzy electrostatic interactions with its disordered C-terminal domain, thereby confining the H3 tail to a wide cavity faced by the Rtt109 active site. These fuzzy interactions between disordered domains achieve localization of the H3 tail to the catalytic site with minimal loss of entropy, and may represent a common mechanism of enzymatic reactions involving highly disordered substrates. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS [1] S. D’Arcy, K. Luger, Curr. Opin. Struct. Biol. 2011, 21(6), 728–734. [2] J. Fillingham, J. Recht, J. F. Greenblatt, Mol. Cell Biol. 2008, 28, 4342–4353. [3] R. ADVERTISERS Driscoll, A. Hudson, S. P. Jackson, Science 2007, 315, 649–652. [4] N. Danilenko, L. Lercher, F. Gabel, J. Kirkpatrick, T. Carlomagno Nature Commun. 2019, 10, AUTHOR INDEX Article number: 3435. PL010 PLENARY LECTURE / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 29 ENRICA BORDIGNON1, NOT ALL MEMBRANE-MIMICKING ENVIRONMENTS LAURA GALAZZO1, ARE THE SAME. AN EPR CASE STUDY FROM IN VITRO SVETLANA KUCHER1, GIANMARCO MEIER2, TO IN CELL. MARKUS SEEGER2 1 Nanobodies ( i.e. single-domain antibodies) are promising new tools for in-cell applications due to their low mo-Ruhr University Bochum, Bochum, Germany. lecular weight, protein- and state- specificity, nano- or sub-nano-molar affinity to their target and the possibility to 2University of Zürich, Zürich, be inserted into cells. We show here how spin-labeled nanobodies can be used as conformational reporters of wild Switzerland type ABC transporters via Double Double Electron Resonance (DEER), a pulsed EPR technique that accurately mea- sures inter-spin distances. First, we show a proof of principle of the use of gadolinium-labeled nanobodies against the heterodimeric exporter TM287/288 [1]. Caveats regarding the nanobody state specificity at micromolar concentrations will be discussed. Second, we present a systematic study on the homodimeric ABC exporter MsbA using non-state-specific nanobodies targeting the two nucleotide binding domains. We gathered structural information in detergent, proteoliposomes, nanodiscs, inside-out vesicles from E. coli and in living cells. We found that there is a remarkable modification of the conformational landscape of the transporter in specific membrane-mimicking environments, proving the need of in-cell structural studies [2]. Advantages and challenges of using biocompatible Gd-labels down to nanomolar concentrations in cells will be also addressed [3]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS [1] Galazzo et al. , PNAS 2020, 117 (5), 2441-2448. [2] Galazzo, Meier, Januliene, De Vecchis, Striednig, Hilbi, Schäfer, Kuprov, Moeller, Bordignon, Seeger, 2021, AUTHOR INDEX in preparation. [3] Kucher et al. , J. Phys. Chem. Lett. 2021, 12, 14, 3679–3684. PL011 PLENARY LECTURE / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 30 IVAN SUDAKOV1,2, PROBING ORGANIC AND HYBRID PEROVSKITE SOLAR MELISSA VAN LANDEGHEM1,2, CELL MATERIALS WITH EPR RUBEN LENAERTS3,4, WOUTER VAN GOMPEL3,4, The constantly growing energy demand and its related negative impact on the environment drives the search for DIRK VANDERZANDE3,4, alternative renewable energy sources. Organic solar cells (OSCs) have great potential for future applications. While WOUTER MAES3,4, fullerene acceptors seemed for a long time unavoidable as acceptor components, non-fullerene acceptors (NFAs) ETIENNE GOOVAERTS2, have recently taken the lead with power conversion efficiencies (PCEs) reaching over 18% for single-junction so-SABINE VAN DOORSLAER1 lar cell design. With these rapid advances in material synthesis and device performance, the long-term stability of 1University of Antwerp, Department of OSCs has become the main remaining obstacle hampering commercialization. In order to overcome this last hur- Chemistry, BIMEF, Antwerp, Belgium 2 dle, a thorough understanding of the photostability of the blend materials in OSCs is needed. In the first part of the University of Antwerp, Department of Physics, NANOrOPT, Antwerp, Belgium talk, the strength of combining different EPR techniques with DFT computations and optical spectroscopy will be 3Hasselt University, Institute for Materials demonstrated in order to analyse the light-induced formation of positive and negative polarons in blend materials Research, Diepenbeek, Belgium 4 and to study the stability of donor and acceptor materials in fullerene-free bulk heterojunction solar cells [1,2]. IMOMEC Division, IMEC, Diepenbeek, Belgium In parallel with the rapid recent rise in the PCEs of OSCs, lead-halide perovskites have been shown to perform excellently as photovoltaic materials. While 3D lead-halide perovskites already abundantly demonstrated their performance in thin-film photovoltaics [3], the 2D hybrid perovskites, consisting of layers of lead-halide octahedra separated by long organic linker cations, have recently emerged as being very promising for functional material design tailored towards specific optical and electronic properties. In the second part of the talk, it will be demonstrated how the earlier mentioned EPR approach can capture the light-induced charge transfer in a series of novel INTRODUCTION 2D perovskites. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] I. Sudakov, M. Van Landeghem, R. Lenaerts, W. Maes, S. Van Doorslaer, E. Goovaerts, Adv. Energy Mater. 2020, 10, 2002095. [2] M. Van AUTHOR INDEX Landeghem, W. Maes, E. Goovaerts, S. Van Doorslaer, J. Magn. Reson. 2018 288, 1-10. [3] H. J. Snaith, J. Phys. Chem. Lett. 2013, 4, 3623-3630. PL012 PLENARY LECTURE / FRONTIERS IN MAGNETIC RESONANCE 31 EUGENIO CORONADO SPIN MOLECULES FOR QUANTUM COMPUTING ICMol-Univ. Valencia, Paterna, Spain; eugenio.coronado@uv.es Spins provide one of the simplest platforms to encode a quantum bit (qubit), the elementary unit of future quantum computers. A challenge in this topic is to control the quantum decoherence in these spin qubits by minimizing the sources of decoherence (dipolar spin-spin interactions, hyperfine interactions and spin-phonon interactions). This loss of quantum information by interaction with the environment can be quantified by the phase memory time T , which for electronic spins can be estimated from pulsed EPR measurements. Here, I will show how a 2 molecular approach can compete with, or even be advantageous to a conventional approach, which is based on extended inorganic lattices. Thus, the versatility of chemistry can be exploited to design robust molecular quantum spin systems showing enhanced decoherences or that host more than one spin qubit in order to implement quantum logic gates [2]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] E. Coronado, Nat. Rev. Mater. 2020, 5, 87-104; [2] A. Gaita-Ariño, F. Luis, S. Hill, E.Coronado, Nature Chem. 2019, 11, 301-309. PL013 PLENARY LECTURE / BIOSOLIDS 32 GUIDO PINTACUDA STRUCTURAL DYNAMICS OF MEMBRANE PROTEINS BY University of Lyon (CNRS/ENS MAGIC-ANGLE SPINNING NMR Lyon/UCBL), High-field NMR Centre, Villeurbanne, France In recent years, magic-angle spinning (MAS) NMR has developed as a powerful technique to investigate structure and dynamics of membrane proteins, enabling the study of these challenging systems in their native-like envi- ronment. Faster (100 kHz and above) MAS rates have paved the way for the direct detection of proton resonances, enabling a boost in sensitivity and resolution with respect to the more traditional approaches. In combination with high magnetic fields, this technical progress revolutionizes the atomic-level investigation of proteins i) by enlarging the molecular size of the systems that can be investigated with site specificity; ii) by reducing the requirements in terms of isotopic labeling, notably deuteration; iii) by speeding up the tedious processes of resonance assignment and acquisition of dynamical parameters. Here we review the strategies underlying this leap forward and describe its potential for the detailed characterization of different transmembrane channels and transporters reconstituted in lipid bilayers. By measuring extended sets of site-specific dynamic observables involving both the backbone and the side chains, we unveil the presence of localized variations in conformational flexibility in regions responsible for substrate selectivity and transport and shed new light on the relation between conformational plasticity and function. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PL014 PLENARY LECTURE / METABOLOMICS 33 ANA M. GIL INCURSIONS OF NMR METABOLOMICS IN DISEASE RESEARCH CICECO-Aveiro Institute of Materials and Department of This talk will cover examples of applications of NMR metabolom- show how NMR metabolomics can help in understanding and, po- Chemistry, University of Aveiro, ics in two different contexts: 1) stem cell (SC) metabolism for guid- tentially, optimizing SC behavior in tissue regeneration medicine. Aveiro, Portugal ance of differentiation onto pure lineages for tissue regeneration Future perspectives to enhance such knowledge will be discussed. improved strategies, and 2) understanding and following-up breast Secondly, this talk will address the process of acquisition of endo-cancer (BC) therapy, in two in vivo murine models of BC (triple-neg- crine therapy resistance in a medroxyprogesterone acetate (MPA) ative (poor prognostic) and endocrine (one of the most common). mouse model of BC, and the use of NMR metabolomics of tumor The first part of this talk is based on SC differentiation as a means for tissue to find resistance metabolic signatures. Implications of the specific tissue regeneration, an ongoing important research field results on therapy schemes for endocrine BC will be briefly dis-with multiple clinical applications. As well as using metabolomics cussed. In addition, metabolomics of a xerograph mouse model of to characterize endo- and exometabolome stepwise adaptations triple-negative BC will also be addressed, including the characteri-during SC differentiation, issues such as inter-donor variability or zation of the tumors themselves upon treatment with cisplatin and SC aging effects are not yet fully known or understood. Here, NMR a novel potential palladium complex, as well as the effects of those cell metabolomics is shown to unveil the full metabolome of adi-drugs on different mouse organs, to assess toxicity effects. The pose tissue SCs and their progression through osteogenesis. Trans- above results exploit the use of different in vivo murine models of versal osteogenesis metabolic markers are suggested, as are appar- BC, to unveil tissue metabolic signatures and their contribution to ent donor-specific metabolites and corresponding pathways. Fur- understanding tumor metabolic dynamics during disease BC pro- thermore, the effects of SC aging, seldomly considered as important gression or therapy. INTRODUCTION underlying contributions, are identified and used to refine global SPONSORS progression markers into osteogenic-only markers. These results PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES Acknowledgements: FCT for funding the BIOIMPLANT project PTDC/BTM-ORG/28835/2017, POCI-01-0145-FEDER-028835; FCT and the Sociedade Portuguesa de Química for grants SFRH/ BD/150655/2020 grant (PhD), SFRH/BD/145920/2019 (PhD); CICECO-Aveiro Institute of Materials project, with references UIDB/50011/2020 & UIDP/50011/2020, financed by national POSTERS funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement; Institute of Biomedicine of Aveiro (iBiMED) with references UID/ ADVERTISERS BIM/04501/2019, POCI-01-0145-FEDER-007628 and pAGE (CENTRO-01-0145-FEDER-000003); the National NMR Network (PTNMR), partially supported by Infrastructure Project № 022161 AUTHOR INDEX (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC) and grant PD/BD/142850/2018 (PhD). PL015 PLENARY LECTURE / ORGANIC AND COMPOSITE SOLIDS 34 LYNDON EMSLEY NEW METHODS IN NMR CRYSTALLOGRAPHY Institut des Sciences et Ingénierie Chimiques, École Structure elucidation of amorphous materials and microcrystalline solids presents one of the key challenges in Polytechnique Fédérale de chemistry today. While techniques such as single crystal diffraction and cryo-electron microscopy are generally Lausanne (EPFL), Lausanne, not able to characterize such materials, we will show how an approach based on measured NMR chemical shifts Switzerland in combination with Machine Learned calculation of shifts from candidate structures can rapidly determine full three-dimensional structures from powders. Further, since the method does not require any significant long-range order, we have recently shown ho it can be extended to determine structures in non-crystalline, amorphous, and hierarchical or composite materials. In the talk we will focus on some of the most recent key methodological advances, which include: • Super-resolution 1H NMR spectroscopy in solids; • Bayesian probabilistic assignment of chemical shifts in organic solids; and • Sensitivity gains provided by dynamic nuclear polarization (DNP). INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX 35 TUTORIAL LECTURE TU001 TUTORIAL LECTURE / FIELD-CYCLING NMR RELAXOMETRY 36 RAINER KIMMICH FIELD-CYCLING NMR RELAXOMETRY: University of Ulm, Ulm, Germany ELUCIDATION OF COMPLEX MOLECULAR DYNAMICS In this tutorial, we will outline the unique strength of field-cycling NMR relaxometry for the characterization of molecular dynamic processes in ordered, macromolecular or porous media [1]. The methodological idea behind and typical experimental set-ups will be described in terms of technical and physical limits at low and high fields. This in particular refers to the Redfield limit, i.e. the condition that the spin-lattice relaxation rates must be less than the fluctuation rates of spin interactions in order to be compatible with the Bloch/Wangsness/Redfield (BWR) relaxation theory. With restricted molecular motions leaving secular spin interactions partially unaveraged, so-called local fields will further limit the experimental range at low fields. Fluctuations of spin interactions, i.e. the thermal phenomena causing spin relaxation, are described and characterized by correlation functions or – via Fourier transform – by spectral densities. We will outline how these functions can be derived for given model scenarios of molecular dynamics in diverse complex systems. The basis of such probability treatments is in each case an adequate equation of motion plus topological constraints imposed by the system. Relevant equations of motion are normal or anomalous rotational or translational diffusion equations and Langevin-type equations predicting distributions of modes in elastically coupled systems. Applications to be discussed in more detail are (i) order director fluctuations in nematic liquid crystals both in bulk and subjected to surface ordering in porous media, (ii) reptation, i.e. the snakelike displacement of polymer seg- INTRODUCTION ments through chain-entangled or pore-confined systems, and (iii) dynamic exchange and displacement processes SPONSORS of fluid adsorbate molecules at surfaces of porous media. The dominating mechanism of the latter scenario will be PROGRAM shown to be “reorientation by translational displacments” (RMTD) along the topology of pore surfaces. This process PROGRAM — BY DAY can be influenced in a non-thermal way by flow-enhanced transport of adsorbate fluids through pore networks. TOPICS Corresponding experimental and computer-simulated data will be presented. Finally, a list of general take-home PRIZE LECTURES rules summarizing this tutorial will be discussed. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] “Field-cycling NMR Relaxometry. Instrumentation, Model Theories and Applications” (Ed. R. Kimmich), Roy. Soc. of Chemistry, AUTHOR INDEX Cambridge, 2019. 37 PARALLEL SESSIONS INVITED AND PROMOTED LECTURES IN001 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULES AND INTERACTIONS I 38 ROLAND RIEK TOWARDS AUTOMATED AND MULTI STATE Laboratory of Physical NMR STRUCTURE DETERMINATION Chemistry, Swiss Federal Institute of Technology, Zürich, Switzerland NMR belongs to one of the main methods for atomic resolution structure determination of biomolecules. In order to automate the 3D structure determination of proteins we implemented machine learning in peak picking and sequential assignment in concert with FLYA and CYANA to fully automatize protein structure determination from amino acid sequence and multi-dimensional NMR spectra (including 2D correlation and 3D backbone triple reso- nance experiments, side chain correlation and NOESY experiments) as input with almost complete success rate on 100 protein systems (with a molecular weight up to ca 20 kDa) tested within 5 hours of calculation time. The exact NOE approach is further discussed as it enables multi-state structure determination of proteins opening along with relaxation measurements an avenue towards a movie of a protein at atomic resolution. Applications therein on 4 different systems ( i.e. PDZ, WW domain, cyclophilin, KRAS) are presented. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN002 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULES AND INTERACTIONS I 39 SAMUEL G. CHAMBERLAIN1, INVESTIGATION OF THE LIPIDATED C-TERMINUS ANDREA GOHLKE2, OF RALA: MUTUALLY EXCLUSIVE BINDING TO DARERCA OWEN1, HELEN R. MOTT1 MEMBRANES OR CALMODULIN. 1Department of Biochemistry, University of Cambridge, Cambridge, U.K. RalA is a small GTPase and a member of the Ras family. This molecular switch is activated downstream of Ras and 2Structure, Biophysics and Fragment-Based is widely implicated in tumour formation and growth. The C-terminus of RalA is isoprenylated, which, along with a Lead Generation, Discovery Sciences, polybasic motif, anchors the protein to the lipid bilayer. We have used nanodiscs to assess the interactions of RalA R&D, AstraZeneca, Cambridge, U.K. with the membrane and found that the bound nucleotide modulates the extent of the interactions, with the inac- tive, GDP-bound RalA binding more tightly to the nanodisc. Previous work has shown that the ubiquitous Ca2+-sensor calmodulin (CaM) binds to small GTPases such as RalA and K-Ras4B but a lack of structural information has obscured the functional consequences of these interactions. We have investigated the binding of CaM to RalA and found that CaM interacts exclusively with the C-terminus of RalA. Biophysical and structural analyses show that the two RalA membrane-targeting motifs (the prenyl anchor and the polybasic motif) are engaged by distinct lobes of CaM and that CaM binding leads to removal of RalA from its membrane environment. The structure of this complex, along with a biophysical investigation into membrane removal, provides a framework with which to under- stand how CaM regulates the function of RalA. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PT001 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULES AND INTERACTIONS I 40 PIOTR GARBACZ1, CROSS-RELAXATION INDUCED BY THE ANTISYMMETRIC PART ANNA ZAWADZKA- OF THE CHEMICAL SHIFT TENSOR KAZIMIERCZUK1, WIKTOR KOŹMIŃSKI1 One of the NMR-based methods of structure determination of mol- is when antisymmetries are parallel to each other, e.g. , most planar 1University of Warsaw, Faculty of ecules, especially those of biological activity such as proteins and molecules fulfill this condition. Chemistry, Warsaw, Poland nucleic acids, is utilizing interferences between nuclear relaxation The dramatic influence of anti-CSA is demonstrated using a com-mechanisms. In particular, the interference due to the dependence bined computational and experimental approach in studies of cis-di-of the magnetic field nearby the nuclei on the orientation of the fluorodichloroethene. It follows from the coupled-cluster quantum molecule with respect to the external magnetic field, e.g. , that which chemical computations performed in the CFOUR computer pro- is caused by the chemical shift anisotropy (CSA), namely the CSA gram that the symmetric parts of 19F nuclei in cis-difluorodichloro-cross-relaxation, may be used for this purpose. For instance, CSA ethene molecule are oriented relative to each other in such a way cross-correlation provides the dihedral angles of the protein back-that almost only anti-CSA contributes to the 19F-19F cross-relaxation bone, facilitating the determination of the overall three-dimension- rate. For this simple model molecule, the magnitude of anti-CSA al structure of the biomolecule [1]. reaches 30 ppm for 19F nuclei, and the contribution of anti-CSA to We show that an overlooked part of the CSA interaction, the so- the total CSA cross-relaxation rate is at least 90%. Our theoretical called vector antisymmetry of chemical shift tensor ( ), defined as results are supported by experimentally determined rate constants /2, contributes to the CSA cross-relaxation rate of 19F-19F cross-relaxation in the 19FCl13C=12C19FCl molecule and mea-constant, and its neglection may cause an erroneous attribution of surements of cross-correlated relaxation constants involving inter-the molecular conformation. action between the CSA and dipole-dipole relaxation mechanisms. INTRODUCTION SPONSORS The rate constant of the CSA cross-correlation caused by the an- Moreover, form the computational studies of amino acids, we found PROGRAM tisymmetric part of CS tensor (anti-CSA) was found using the that anti-CSAs of 15N/13C nuclei are of the order of several ppm. a PROGRAM — BY DAY Bloch-Redfield-Wangsness theoretical description of nuclear re- Therefore, the anti-CSA contribution to CS-induced cross-correla- TOPICS laxation processes [2]. We found that this rate constant for the tion affects the structure determination of an oligopeptide. Conse- PRIZE LECTURES cross-correlation of anti-CSA of nuclei I and S is proportional to the quently, the proper analysis of molecular conformation using CSA PLENARY LECTURES factor , where is the gyromagnetic ratio of the cross-correlation requires introducing a new component that de- TUTORIAL LECTURE nucleus I/ S and B is the strength of the magnetic field. Therefore, pends on the antisymmetric part of the chemical shift tensor. INVITED AND PROMOTED LECTURES the most favorable orientation of anti-CSA represented as a vector POSTERS ADVERTISERS References: [1] B. Reif, A. Diener, M. Hennig, M. Maurer, C. Griesinger, J. Magn. Reson. 2000, 143, 45–68. [2] M. Goldman, J. Magn. Reson. 2001, 149, 160–187. AUTHOR INDEX Acknowledgments: PG acknowledges the National Science Centre, Poland, for the financial support through OPUS 16 Grant No. 2018/31/B/ST4/02570. PT002 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULES AND INTERACTIONS I 41 QIANG LI1,2, 4’-F URIDINE-A SENSITIVE LABEL FOR PROBING RNA STRUCTURE MARKO TRAJKOVSKI2, AND FUNCTION BY 19F NMR JANEZ PLAVEC2, CHUANZHENG ZHOU1 Due to the high sensitivity in chemical shift, the 100% natural Herein, we reported a more mature strategy through a selective pro-1Nankai University, College of abundance and the pronounced bioorthogonality of 19F, 19F NMR has tection of the hydroxy groups in stages to synthesize 4’-F-modified Chemistry, State Key Laboratory of been successfully used to investigate secondary structure and func- uridine phosphoramidite monomers, which was stable enough to Elemento-Organic Chemistry and Department of Chemical Biology, tion of nucleic acids for more than a decade [1]. There are several then be incorporated into longer oligonucleotides through stan-Tianjin, China main strategies in the field of fluorine-labeling, one of which is the dard solid-phase synthesis. Therfore, we incorporated U4’-F into oli-2National Institute of Chemistry, introduction of fluorine(s) at sugar, usually C2’-position [2]. Among gos and investigated their biophysical and biochemical properties. Slovenian NMR Centre, Ljubljana, Slovenia them, 2’-F-labeled RNA is powerful tool, but it directly affect ribose The 4’-F modification does not significantly alter the nature of RNA puckering [2]. and therefore can mimic the state of native RNA for structural stud- ies. 19F NMR results show that this modification can not only distin- Structurally, C4’-position should be an attractive modification site guish between ssRNA and dsRNA, but also identify mismatches and in 19F-labeling. First of all, C4’-substitutions locate on the edge of the binding of RNA-processing proteins with chemical shift dispersion minor groove upon duplex formation, with no significantly affect- up to 4 ppm, indicating that this modification can be widely used ing the helical conformation [3]. Secondly, C4’-position is a member through NMR spectroscopy for determination various RNA struc- of the phosphate backbone (P -O5’-C5’-C4’-C3’-O3’-P ) which has di- 0 -1 tures [5]. rectly interactions with proteins, such as cGAS and ribonucleases. Unfortunately, due to the instability of 4’-F-modified nucleosides, scientists have not successfully introduced them into the RNA INTRODUCTION strands [4]. The phenomenon, cleavage of glycosidic bonds in con- SPONSORS ventional experimental solutions, is the main reason for limiting its PROGRAM continued research. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] H. Chen, L. Peng, et. al, Chem Soc Rev 2013, 42, 7971-7982. [2] K. Christoph, M. Ronald, J Am Chem Soc 2005, 127, 1158-11559. [3] D. Summerer, A. Marx, J Am Chem Soc 2002, POSTERS 124, 910-911. [4] L. Seongmi, V. Gregory, Org Lett 2007, 9, 5007-5009. [5] Q. Li, J. Plavec, C. Zhou, J Am Chem Soc 2020, 142, 4739-4748 ADVERTISERS Acknowledgements: This work was supported by the National Natural Science Foundation of China (Grant Nos. 91953115, 21877064, and 21740002) and by the Slovenian Research Agency AUTHOR INDEX (ARRS, Grant Nos. P1-0242 and J1-1704). PT003 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULES AND INTERACTIONS I 42 SIRINE NOURI1, BACTERIAL TYPE 1 PILI AS A NEW VERSATILE ALIGNMENT JULIEN BOUDET2, MEDIUM FOR NMR SPECTROSCOPY HIANG DREHER-TEO2, FRÉDÉRIC H.-T. ALLAIN3, The measurement of NMR Residual Dipolar Couplings (RDCs) is a allows for efficient NMR sample preparation. In agreement with the LOÏC SALMON1, powerful tool to gain information on atomic-level structure and high stability of the pilus against denaturation, the alignment me-CHRISTOPH GIESE2 dynamics of biomolecules. RDCs become measurable when a mol- dium was stable at all temperature tested (4 - 50°C) and withstood 1Centre de RMN à Très Hauts Champs, ecule is partially aligned with respect to the NMR magnetic field. To challenging conditions such as pH 1.5, the presence of commonly University of Lyon, Villeurbanne, reach this partial alignment, a variety of anisotropic media such as used detergents and lipids (including 1% (w/v) SDS), or 50% (v/v) France 2Institute of Molecular Biology and bicelles, strained gels or filamentous phages can be used to induce DMSO, DMF or methanol. To demonstrate the applicability of the Biophysics, ETH Zurich, Zurich, predominantly steric or electrostatic alignment [1,2]. However, type 1 pilus alignment medium, we measured RDCs of three differ-Switzerland 3 finding suitable alignment media compatible with various types of ent systems - a protein (human ubiquitin), an RNA (HIV-1 TAR) and a Institute of Biochemistry, ETH Zurich, Zurich, Switzerland molecules and a broad range of experimental conditions remains a small organic molecule (camphor) – and show that in each case, ex-challenge. perimental RDCs were highly correlated with RDC values back-cal- culated from high-resolution structures of these molecules. Overall Here we present a new liquid-crystalline alignment medium com- the alignment induced by type 1 pili appears to arise mainly from posed of purified type 1 pilus rods from Escherichia coli, which are steric interactions with a negative electrostatic contribution. filamentous protein polymers extending from the surface of the cell into the extracellular space [3,4]. Remarkably, under physiological In summary, the pilus-based medium is compatible with challeng-conditions type 1 pilus rods have practically infinite kinetic stabil- ing experimental conditions and is usable with both biological ity against spontaneous dissociation/unfolding [5]. Using a recom- macromolecules and small organic molecules. These results sug- INTRODUCTION binant system to elongate the wild-type pilus rod from an average gest that type 1 pili should represent a very useful complement to SPONSORS length of ~670 nm to ~2100 nm, we show that alignment is achieved existing alignment media and may assist molecular structure or PROGRAM at a minimum pilus concentration of ~15 mg/ml, which togeth- dynamics determination by NMR. PROGRAM — BY DAY er with a yield of ~17 mg pili obtained per liter of bacterial culture TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: INVITED AND PROMOTED LECTURES [1] M. Blackledge, Prog. Nucl. Magn. Reson. Spectrosc, 2005, 46, 23-61. [2] J.R. Tolman, K. Ruan, Chem. Rev. 2006, 106, 1720-1736. POSTERS [3] E. Hahn, P. Wild, U. Hermanns, P. Sebbel, R. Glockshuber, M. Haner, N. Taschner, P. Burkhard, U. Aebi, S.A. Muller, J. Mol. Biol. 2002, 323, 845-857. ADVERTISERS [4] C. C. Brinton Jr, Trans. N. Y. Acad. Sci. 1965, 27, 1003-1054. AUTHOR INDEX [5] C. Puorger, O. Eidam, G. Capitani, D. Erilov, M.G. Grutter, R. Glockshuber. Structure 2008, 16 , 631-42. IN003 PARALLEL SESSIONS / INVITED SPEAKER / NMR OF QUANTUM MATERIALS 43 MLADEN HORVATIĆ1, DISTINGUISHING THE DIMENSIONALITY OF A SPIN MARTIN KLANJŠEK2, SYSTEM: DETERMINATION OF THE TOMONAGA- EDMOND ORIGNAC3, RANJITH KUMAR LUTTINGER PARAMETER K IN QUASI-1D SYSTEMS KIZHAKE MALAYIL1 1 The nuclear magnetic resonance (NMR) in antiferromagnetic quantum spin materials has recently provided a ma- Laboratoire National des Champs Magnétiques Intenses, jor result regarding quasi-one-dimensional (quasi-1D) systems, namely an important improvement of the standard, LNCMI-CNRS, EMFL, “Tomonaga-Luttinger liquid” (TLL), low-energy description of these systems in their gapless regime (typically in-Université Grenoble Alpes, UPS duced by magnetic field). This purely 1D description is characterized by power-law dependence of the response/ and INSA Toulouse, Grenoble, France correlation functions, whose exponents are defined by the TLL parameter K. However, at low temperature the ef-2Jožef Stefan Institute, Ljubljana, fects of 3D exchange couplings significantly modify the TLL response, which can be theoretically described by a Slovenia 3Université de Lyon, ENS de random phase approximation (RPA). We confirm the validity of the recently derived RPA correction for the spin- Lyon, Lab. de Physique, UCBL, spin correlation functions [1], by applying the corresponding formula to the temperature dependence of the NMR CNRS, Lyon, France relaxation rate T -1 in the two representative quasi-1D spin compounds, (C H N) CuBr (DIMPY) and BaCo V O . A 1 7 10 2 4 2 2 8 successful fit of the T -1 data provides the first direct experimental determination of the K values that perfectly agree 1 with the theoretically calculated ones [2]. In general, by taking into account the RPA correction, we strongly broaden the scope of the TLL description, which becomes applicable to less quasi-1D compounds/systems approaching the 3D regime. The RPA correction can be easily calculated for any geometry of the 3D exchange couplings, and provides a T -1( T) dependence that is clearly 1 INTRODUCTION different from those observed in either quasi-2D [3] or in 3D spin systems. NMR T -1 data thus provide an exception-1 SPONSORS ally simple way to distinguish the effective dimensionality of a gapless spin system. In quasi-1D systems, from T -1 1 PROGRAM data we directly determine the K parameter, providing insight into the nature and the strength of the quasiparticle PROGRAM — BY DAY interactions. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] M. Dupont et al., Phys. Rev. B. 2018, 98, 094403/1-16. [2] M. Horvatić, M. Klanjšek, E. Orignac, Phys. Rev. B. 2020, 101, 220406(R)/1-5. [3] K. M. AUTHOR INDEX Ranjith et al. , unpublished. IN004 PARALLEL SESSIONS / INVITED SPEAKER / NMR OF QUANTUM MATERIALS 44 ANDREJ ZORKO1, 2 INTRIGUING SPINON-METAL STATE OF AN INSULATING 1Jožef Stefan Institute, Ljubljana, KAGOME LATTICE ANTIFERROMAGNET Slovenia 2Faculty of Mathematics and Physics, University of Ljubljana, Spinon metal is a fascinating state of mater found in certain charge insulators that feature strong geometrical frus-Ljubljana, Slovenia tration of the underlying spin lattice. Contrary to more common ferromagnetically or antiferromagnetically or- dered states, it belongs to a class of states known as quantum spin liquids. These highly quantum entangled states remain magnetically disordered down to zero temperature due to strong quantum fluctuations and feature exotic magnetic excitations known as spinons. In the case of a spinon metal, spinons form a Fermi surface and may, in a way, behave similarly as itinerant electrons behave in ordinary metals. Realizations of spinon metals in real materials are extremely rare. I will present such case that has been recently confirmed in the kagome lattice antiferromagnet Zn-brochantite, ZnCu (OH) SO , via local-probe techniques in-3 6 4 cluding muon spectroscopy and nuclear magnetic resonance. Quite unexpectedly, these investigations have re- vealed that the spinon-metal state in Zn-brochantite undergoes a magnetic-field induced instability due to spinon pairing at low temperatures [1]. This phenomenon is analogous to the formation of Cooper pairs of electrons in superconductors. Moreover, another phenomenon traditionally associated with itinerant electrons has been recently discovered in this material [2] in which the charge degrees of freedom are frozen. Namely, magnetic moments of impurities present in Zn-brochantite become screened by spinons at low temperatures in an event that resembles Kondo screening of magnetic impurities by conduction electrons in metals. In this spinon Kondo effect that was theoretically predicted a while ago but has been observed experimentally for the first time in Zn-brochantite [2], INTRODUCTION the role of electrons in Kondo screening is taken over by chargeless spinons. Both phenomena, spinon pairing and SPONSORS spinon Kondo effect, make spinon metals highly attractive for emerging quantum technologies. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] M. Gomilšek, M. Klanjšek, R. Žitko, M. Pregelj, F. Bert, P. Mendels, Y. Li, Q. M. Zhang, and A. Zorko, Phys. Rev. Lett. 2017, 119, 137205. [2] M. POSTERS Gomilšek, R. Žitko, M. Klanjšek, M. Pregelj, C. Baines, Y. Li, Q. M. Zhang, and A. Zorko, Nat. Phys. 2019, 15, 754-758. ADVERTISERS Acknowledgements: The financial support of the Slovenian Research Agency through the Program No. P1-0125 and Projects No. N1-0148 and J1-2461 is AUTHOR INDEX acknowledged. PT004 PARALLEL SESSIONS / PROMOTED TALK / NMR OF QUANTUM MATERIALS 45 TAKASHI IMAI INVERSE LAPLACE TRANSFORM (ILT) T ANALYSIS 1 Department of Physics and AS A NEW PROBE OF QUANTUM MATERIALS WITH Astronomy, McMaster University, Hamilton, Canada DISORDER Magnetic materials generally undergo a phase transition into a magnetically ordered ground state at low temperatures. When the spin-spin interactions are geometrically frustrated, such a phase transition may be prevented and a novel quantum spin liquid (QSL) sate may be realized instead, in which spins are highly entangled but remain paramagnetic [1]. Examples of such QSL’s include Kitaev lattice formed by Ising spins arranged on the honeycomb structure, and spins interacting with Heisenberg’s exchange interaction on the the kagome lattice ( i.e. corner-sharing triangular lattice). Identifying such a QSL material is a holy grail of today’s quantum condensed matter physics, and NMR techniques plays a pivotal role in characterizing their local magnetic properties [2]. A common trait shared by various QSL candidate materials is that their ground state properties are sensitive to structural disorder, which makes proper characterization of the magnetic ground state difficult due to the distribution of the NMR parameters. In this talk, we will demonstrate how one could overcome this difficulty by deducing the density distribution function P(1/T ) of the NMR spin-lattice relaxation rate using the inverse Laplace transform techniques [3-6]. 1 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] T. Imai and Y. S. Lee, Physics Today 2016, 69, 30-36. [2] M. Fu et al., Science 2015, 350, 655-658. [3] P. M. Singer et al., Phys. Rev. B 2020, 101, AUTHOR INDEX 174508. [4] S.K. Takahashi et al. Phys. Rev. X 2019, 9, 031047. [5] J. Wang et al., Phys. Rev. B 2021, in press. [6] J. Wang et al., 2021, in press. PT005 PARALLEL SESSIONS / PROMOTED TALK / NMR OF QUANTUM MATERIALS 46 SERGEI ZVYAGIN PRESSURE-TUNED MAGNETIC INTERACTIONS Dresden High Magnetic Field IN A TRIANGULAR-LATTICE QUANTUM Laboratory (HLD), Helmholtz- Zentrum Dresden Rossendorf ANTIFERROMAGNET (HZDR), Dresden, Germany Quantum triangular-lattice antiferromagnets are important prototype systems to investigate numerous phenome- na of the geometrical frustration in condensed matter. Apart from highly unusual magnetic properties, they possess a rich phase diagram (ranging from an unfrustrated square lattice to a quantum spin liquid), yet to be confirmed experimentally. One major obstacle in this area of research is the lack of materials with appropriate (ideally tuned) magnetic parameters. Using Cs CuCl as a model system, we demonstrate an alternative approach, where, instead 2 4 of the chemical composition, the spin Hamiltonian is altered by hydrostatic pressure [1]. The approach combines high-pressure electron spin resonance and r.f. susceptibility measurements, allowing us not only to quasi-continuously tune the exchange parameters, but also to accurately monitor them. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] S.A. Zvyagin, D. Graf, T. Sakurai, S. Kimura, H. Nojiri, J. Wosnitza, H. Ohta, T. Ono, and H. Tanaka, Nat. Comm. 2019, 10, 1064. TUTORIAL LECTURE Acknowledgements: This work was partially supported by the Deutsche Forschungsgemeinschaft (projects ZV6/2-2, SFB 1143, and through the Würzburg- INVITED AND PROMOTED LECTURES Dresden Cluster of Excellence on Complexity and Topology in Quantum Matter - ct.qmat (EXC 2147, project-id 39085490)). We acknowledge support by the HLD at HZDR, member of the European Magnetic Field Laboratory (EMFL). ESR experiments were performed at the High Field Laboratory for Superconducting POSTERS Materials, Institute for Material Research, Tohoku University (proposals 18H0059 and 17H0071). H.N. acknowledges the support of the KAKENHI 16H04005 ADVERTISERS Program. S.Z. acknowledges the support of the ICC-IMR Visitor Program at the Tohoku University. A portion of this work was performed at the National AUTHOR INDEX High Magnetic Field Laboratory, which is supported by the National Science Foundation Cooperative Agreement No. DMR-1644779 and the State of Florida. PT006 PARALLEL SESSIONS / PROMOTED TALK / NMR OF QUANTUM MATERIALS 47 75 ŽIGA GOSAR1, 2, AS NQR STUDY OF QUASI ONE-DIMENSIONAL BING LV3, A MO AS SUPERCONDUCTORS DENIS ARČON1, 2 2 3 3 1Institute Jožef Stefan, Ljubljana, A Mo As , where A stands for K, Rb or Cs, is a family of quasi one-dimensional conductors, where MoAs form quasi 2 3 3 Slovenia 2 one-dimensional double-walled subnanotubes, with A ions in between chains. It is isostructural to the A Cr As Faculty of Mathematics and 2 3 3 Physics, University of Ljubljana, family, which was shown to be superconducting at low temperatures, with [1], [2] and Ljubljana, Slovenia [3], for A = K, Rb and Cs respectivelly. The Molybdenum family achieves higher critical temperatures of 3The University of Texas at Dallas, [4], [5] and [6] for A = K, Rb and Cs respectivelly. Richardson, Texas, USA In this talk, a comprehensive 75As NMR study of of A Mo As for A= K, Rb, Cs is presented [5]. The 75As NQR spectra 2 3 3 show surprising results, indicating four NQR lines, not conforming to the generally accepted structure and space group ( ), which suggests just two chemically inequivalent As sites. Recent density functional theory and neu- tron total scattering studies of K Cr As propose structural instability in the original structure and a local deforma-2 3 3 tion to a space group Amm2 [7]. A possibility of a similar occurance in K Mo As is thus explored here. 2 3 3 Both A Cr As and A Mo As show a power-law temperature dependencies of spin-lattice relxation rates, which are 2 3 3 2 3 3 fingerprints of the Tommonaga-Luttinger liquid behaviour above . However, there is an important difference between the two families of materials with A Cr As showing repulsive interactions and Rb Mo As showing attrac-2 3 3 2 3 3 tive interactions. Interestingly, 75As NQR and 87Rb NMR show different spin-lattice relaxation rate dependence with temperature in Rb Mo As . In the K Mo As , even the different As sites show different spin-lattice relaxation rates, 2 3 3 2 3 3 INTRODUCTION suggesting the importance of multi-orbital physics. SPONSORS The importance of multi-orbital physics is again shown in the superconducting state. No Hebel-Slichter cohrenece PROGRAM peak was observed in both Rb Mo As and K Mo As , the first sign of unconventional superconductivity. 75As NQR in 2 3 3 2 3 3 PROGRAM — BY DAY Rb Mo As shows BCS gap, while 87Rb NMR shows a reduced gap. In the K Mo As again, even different As sites show 2 3 3 2 3 3 TOPICS different behaviour in the superconducting state, with one site observing the BCS gap, and the others following the PRIZE LECTURES Korringa relation even below . PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] J.-K. Bao, et al, Phys. Rev. X. 2015, 5, 011013; [2] J. Yang, et al, PRL 2015, 115, 147002; [3] Z.-T. Tang, et al, Sci China Mater. 2015, 58: 16–20; [4] Q.-G. ADVERTISERS Mu, et al, Science Bulletin. 2018, 63, 952-956; [5] Ž. Gosar, et al, PRB 2020, 101, 220508(R); [6] K. Zhao, et al, APL Mater. 2020, 8, 031103; [7] K. M. Taddei, et al, AUTHOR INDEX PRL 2018, 121, 187002. IN005 PARALLEL SESSIONS / INVITED SPEAKER / IN-CELL MAGNETIC RESONANCE 48 MARC BALDUS CELLULAR SOLID-STATE NMR SPECTROSCOPY: NMR Spectroscopy, Bijvoet RECENT PROGRESS AND APPLICATIONS Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands Solid-state NMR (ssNMR) spectroscopy provides increasing pos- In our presentation, recent methodological progress will be dis- sibilities to study biomolecules in their native pro– or eukaryotic cussed. In addition, we report on applications in the context of cellular setting. To conduct such studies, we have designed pre-whole-cell biocatalysis [8] and the study of protein-protein in- parative procedures that reduce unwanted background labelling teractions in bacterial and human cells. In particular, we demon-by dedicated expression [1,2], purification [3] and protein delivery strate the use of cellular ssNMR to study the supramolecular struc- [4] methods. In addition, we have addressed challenges regarding ture and dynamics of the lipoprotein BamC as part of the β-barrel spectroscopic sensitivity by either using 1H-detection (see, e.g. [5]) or assembly machinery (BAM) complex that catalyzes -barrel protein Dynamic Nuclear Polarization (DNP) to enhance spectroscopic sig- insertion into the outer membrane of E.coli (see, e.g. [8,9]). Using nals. In the latter case, we have investigated the use of hydrophilic 1H ssNMR we have studied BAM complexes containing BamC in tritryl-nitroxides together with Liu et al. [7] because of their favour-lipid bilayers and cellular environments. This method allowed able magnetic-field strength dependence. us to probe the structural and dynamic changes of BamC in dif- ferent protein complexes and membrane environments on a res- idue-specific level. Our results reveal a remarkable topological variability of BamC, which may be critical during different stages of the BAM-mediated substrate insertion process in bacteria [10]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] S. Narasimhan, C. Pinto, A. Lucini Paioni, J. van der Zwan, G. E. Folkers, M. Baldus, Nature Prot. , 2021 16, 893-918. [2] L. A. Baker, M. Daniels, E. A. W. van der Cruijsen, G. E. TUTORIAL LECTURE Folkers, M. Baldus, J. Biomol. NMR, 2015 62, 199-208. [3] Y. Luo, S. Xiang, P. J. Hooikaas, L. van Bezouwen, A. S. Jijumon, C. Janke, F. Förster, A. Akhmanova, M. Baldus, Nature Comm., 2020 INVITED AND PROMOTED LECTURES 11, 18. [4] S. Narasimhan, S. Scherpe, A. Lucini Paioni, J. van der Zwan, G. E. Folkers, H. Ovaa, M. Baldus, Angew Chem Int Ed Engl, 2019, 58, 12969-12973 (2019).[5] L. A. Baker, T. Sinnige, P. Schellenberger, J. de Keyzer, C. A. Siebert, A. J. M. Driessen, M. Baldus, K. Grunewald, Structure 2018, 26, 161-170. [6] Zhai, A. Lucini Paioni, X. Cai, S. Narasimhan, J. Medeiros-Silva, W. Zhang, POSTERS A. Rockenbauer, M. Weingarth, Y. Song, M. Baldus, Y. Liu, J. Phys Chem.B, 2020, 124, 9047-9060. [7] S. Chordia, S. Narasimhan, A. Lucini Paioni, M. Baldus, G. Roelfes, Angew Chem Int Ed Engl, ADVERTISERS 2021 60, 5913-5920. [8] C. Pinto, D. Mance, T. Sinnige, M. Daniels, M. Weingarth, M. Baldus,, Nature Comm., 2018, 9. [9] C. Pinto, D. Mance, M. Julien, M. Daniels, M. Weingarth, M. Baldus, J. AUTHOR INDEX Struct. Biol. , 2019, 206, 1-11.[10] S Xiang, C Pinto, B. Peronne, M. Baldus 2021, in preparation. IN006 PARALLEL SESSIONS / INVITED SPEAKER / IN-CELL MAGNETIC RESONANCE 49 ANNALISA PIERRO1, PROBING PROTEIN STRUCTURAL DYNAMICS IN-CELL ALESSIO BONUCCI1, BY NITROXIDE-BASED SDSL-EPR KETTY C. TAMBURRINI2, AXEL MAGALON3, Understanding how the intracellular medium modulates protein structural dynamics and protein-protein interac- BARBARA ZAMBELLI4, tions is an intriguing but required topic scientists search to address by studying biomolecules in their native envi-FRANÇOIS VIBERT5, ronment. As the cellular environment can not be reproduced in vitro, investigation of biomolecules directly inside BRUNO GUIGLIARELLI1, cells has attracted a growing interest in the past decade. Indeed, efforts in magnetic resonance spectroscopies have GUILLAUME GERBAUD1, enabled important improvements in the study of structural dynamics directly in the cellular context. EMILIEN ETIENNE1, OLIVIER OUARI5, Among magnetic resonances approaches, site-directed spin labeling coupled to electron paramagnetic resonance VALÉRIE BELLE1, spectroscopy (SDSL-EPR) has demonstrated to be one of the powerful approaches to study structural properties of ELISABETTA MILEO1 biomolecules[1,2]. In particular, nitroxide-based SDSL-EPR couples the benefits of high sensitivity and the lack of 1 size constraints for the biomolecule of interest with the ability to study protein structural transitions and interac-Aix Marseille Univ, CNRS, BIP, Marseille, France 2 Aix Marseille Univ, CNRS, AFMB, Marseille, France tions at physiological temperature. 3 Aix Marseille Univ, CNRS, LCB, Marseille, France 4 Laboratory of Bio-Inorganic Chemistry, University In this talk, we will discuss the results achieved in the investigation of the structural dynamics features of several of Bologna, Bologna, Italy cytosolic proteins directly inside cells, by combining the use of nitroxide labels and EPR (cw-EPR and pulsed dipolar 5 Aix Marseille Univ, CNRS, ICR, Marseille, France experiments) spectroscopy. Furthermore, we will focus on NarJ protein [3], for which, beside reaveling its structural features directly in E. coli, we verify its activity inside cells, by evaluating its ability in restoring the activity of its biological partner, the nitrate reductase. INTRODUCTION These results represents a step forward in the development of EPR-based cellular structural biology that goes be- SPONSORS yond the feasibility and opens new perspectives to address biological questions directly inside the cell. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] A. Bonucci, O. Ouari, B. Guigliarelli, V. Belle, E. Mileo, ChemBioChem 2020, 21, 451-460. [2] F. Torricella, A. Pierro, E. Mileo, V. Belle, A. Bonucci, ADVERTISERS BBA-Proteins and Proteomics 2021, 1869, 140653. [3] G. Karthikeyan, A. Bonucci, G. Casano, G. Gerbaud, S. Abel,V. Thomé, L. Kodjabachian, A. Magalon, B. AUTHOR INDEX Guigliarelli, V. Belle,O. Ouari, E. Mileo, Angew. Chem. Int. Ed. 2018, 130, 1380-1384. IN007 PARALLEL SESSIONS / INVITED SPEAKER / IN-CELL MAGNETIC RESONANCE 50 ENRICO LUCHINAT1,2, PROTEIN-DRUG INTERACTIONS MONITORED BY LETIZIA BARBIERI1,3, TIME-RESOLVED NMR IN HUMAN CELLS MATTEO CREMONINI1,4, MATTEO PENNESTRI5, In-cell NMR can investigate protein conformational changes and chemical modifications at atomic resolution di- ALESSIO NOCENTINI2, rectly in living cells. The approach has recently shown great potential in the context of drug development, as it CLAUDIU T. SUPURAN2, can investigate directly the interaction between a ligand and its target protein in its physiological environment. As LUCIA BANCI1,4 such, in-cell NMR can provide precious insights on the intracellular drug-target interaction that can be beneficial 1CERM – Magnetic Resonance Center, to assess the efficacy of active compounds at an early stage of drug development and to increase the chances of suc-University of Florence, Sesto Fiorentino, Italy 2 cess in the subsequent pre-clinical and clinical studies. Furthermore, NMR bioreactors can greatly improve the cell Neurofarba Department, University of Florence, Sesto Fiorentino, Italy sample stability over time and allow monitoring the evolution of intracellular processes, including protein-drug 3CIRMMP – Consorzio Interuniversitario interactions, by time-resolved in-cell NMR. Here, the latest developments of in-cell NMR applied to protein-drug in-Risonanze Magnetiche di Metalloproteine, teractions in human cells are overviewed. First, drug screening in human cells is applied to investigate a set of novel Sesto Fiorentino, Italy 4Department of Chemistry, University of carbonic anhydrase (CA) inhibitors, which are found to bind the intracellular target in a dose- and time-dependent Florence, Sesto Fiorentino, Italy manner [1]. The same approach is applied to study the off-target binding to CA of drugs originally developed to 5Pharmaceutical Business Unit, Bruker UK Limited, Coventry, United Kingdom interact with other targets [2]. Then, an optimized design of in-cell NMR bioreactor is applied to measure drug diffusion and binding in real time [3,4]. Finally, a novel application of in-cell NMR is reported, where intracellular ligand binding affinities are measured in human cells by competition binding experiments. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] E. Luchinat, L. Barbieri, M. Cremonini, A. Nocentini, C. T. Supuran, L. Banci, Angew. Chem. Int. Ed. Engl. 2020, 59, 6535–6539. [2] E. Luchinat, ADVERTISERS L. Barbieri, M. Cremonini, A. Nocentini, C. T. Supuran, L. Banci, ACS Chem Biol 2020, 15, 2792–2800. [3] E. Luchinat, L. Barbieri, T. F. Campbell, L. Banci, Anal. AUTHOR INDEX Chem. 2020, 92, 9997–10006. [4] L. Barbieri, E. Luchinat, JoVE (Journal of Visualized Experiments) 2021, e62323. IN008 PARALLEL SESSIONS / INVITED SPEAKER / IN-CELL MAGNETIC RESONANCE 51 OLAV SCHIEMANN TRITYL SPIN LABELS: Institute of Physical and STUDYING BIOMOLECULAR CONFORMATIONS WITHIN Theoretical Chemistry, University of Bonn, Bonn, Germany. CELLS BY PULSED DIPOLAR EPR The understanding of biomolecular function is coupled to knowledge about the structure and dynamics of these biomolecules, preferably acquired under native conditions. In this regard, pulsed dipolar EPR spectroscopy (PDS) in conjunction with site–directed spin labeling (SDSL) is an important method in the toolbox of biophysical chemistry. However, the currently available spin labels have diverse deficiencies for in–cell applications, for example, low radical stability or long bioconjugation linkers. In the talk, synthesis strategies are introduced for the derivatization of trityl radicals with a maleimide–group, which enables bioconjugation of the trityl labels to cysteins [1-4]. Especially, the resulting trityl spin labels SLIM [3] and oxSLIM [4] yield narrow distance distributions, enable highly sensitive distance measurements down to concentrations of 45 nM, and show high stability against in cell reduction. Using these labels in combination with the Double Quantum Coherence experiment (DQC), the guanine–nucleotide dissociation inhibitor (GDI) domain of Yersinia outer protein O (YopO) is shown to adopt in eukariotic cells a different conformational ensemble than in vitro [3]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES [1] J.J. Jassoy, A. Berndhäuser, F. Duthie, S.P. Kühn, G. Hagelueken, O. Schiemann Angew. Chem. Int. Ed. 2017, 56, 177-181. [2] J.J. Jassoy, C.A. Heubach, T. Hett, POSTERS F. Bernhard, F.R. Haege, G. Hagelueken, O. Schiemann Molecules 2019, 24, 2735. [3] N. Fleck, C.A. Heubach, T. Hett, F.R. Haege, P.P. Bawol, H. Baltruschat, O. ADVERTISERS Schiemann Angew. Chem. Int. Ed. 2020, 59, 9767-9772. [4] N. Fleck, C. Heubach, T. Hett, S. Spicher, S. Grimme, O. Schiemann Chem. Eur. J. 2021, 27, 5292-5297. AUTHOR INDEX Acknowledgements: Funding by the DFG via SPP1601 is gratefully acknowledged. PT007 PARALLEL SESSIONS / PROMOTED TALK / IN-CELL MAGNETIC RESONANCE 52 SHARI L. MEICHSNER1, IN-CELL CHARACTERIZATION OF THE STABLE YURY KUTIN1, TYROSYL RADICAL IN E. COLI MÜGE KASANMASCHEFF1 1Department of Chemistry and RIBONUCLEOTIDE REDUCTASE VIA Chemical Biology, Technische Universität Dortmund, Dortmund, ADVANCED EPR SPECTROSCOPY Germany Determining the structure of biomolecules in their intracellular en- similar to those of Y • in vitro. Next, we performed orientation-se- 122 vironment is fundamental to understand their function. Yet, achiev- lective electron-nuclear double resonance (ENDOR) spectroscopy at ing high resolutions within the cell possesses a great challenge. In 34 GHz to probe the H-bonding environment of Y • in living cells. 122 this work, we employed advanced electron paramagnetic resonance The analysis of our ENDOR data with a significantly high signal-to- (EPR) spectroscopic methods to characterize the structure and dy- noise ratio displayed that the number of hydrogens coupled to Y • 122 namics of Y •, the stable tyrosyl radical in E. coli ribonucleotide in the cells and in vitro is the same. At last, we detected distances 122 reductase (RNR) in whole cells at atomic resolution. between two Y •s residing in each 122 β monomer of E. coli RNR via double electron-electron resonance (DEER). These orientation-se- The E. coli RNR, a paradigm for class Ia enzymes including human lective experiments revealed insights into not only the in vivo struc-RNR, catalyzes the biosynthesis of DNA building blocks and requires ture and dynamics of Y • and β2 subunit but also the in vivo radical a di-iron tyrosyl radical (Y •) cofactor for activity.[1] The knowl- 122 122 distribution within β2.[2] edge on in vitro Y • structure and its radical distribution within 122 β2 subunit has accumulated over the years; yet, little information Additionally, we have site-specifically incorporated 2,3,5-trifluo- exists on in vivo Y •. Therefore, determining in vivo structure and rotyrosine (F Y) at residue 122, generated and identified its radical 122 3 INTRODUCTION distribution of Y • in RNR is essential to understand the function form F Y • in the cells, and obtained in vivo distances between 122 3 122 SPONSORS of the protein in its native intracellular environment. First, we em- F Y •s. This marks the first spectroscopic verification of the gen- 3 122 PROGRAM ployed multi-frequency EPR (9, 34, and 94 GHz) spectroscopy to eration of an unnatural amino acid radical in whole cells providing PROGRAM — BY DAY characterize the generated Y • radical in whole E. coli cells. Our ex- a new possibility for investigating the structure and role of tyrosyl 122 TOPICS perimental data combined with spectral simulations demonstrated radicals involved in fundamental processes such as photosynthesis, PRIZE LECTURES that the structure and environment of Y • in the cells are highly reduction of O to water, and DNA repair in living cells. 122 2 PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] U. Uhlin, H. Eklund, Nature, 1994, 370,533-539. [2] S.L. Meichsner, Y. Kutin, M. Kasanmascheff, Angew. Chem. Int. Ed. , 10.1002/anie.202102914. PT008 PARALLEL SESSIONS / PROMOTED TALK / IN-CELL MAGNETIC RESONANCE 53 SERGEY OVCHERENKO1, 2, UPTAKE OF RL2 BY HUMAN LUNG CANCER CELLS: OLGA CHINAK3, MONITORING BY EPR AND CONFOCAL MICROSCOPY ANTON CHECHUSHKOV3, SERGEY DOBRYNIN1, A recombinant κ-casein fragment – intrinsically disordered recombinant lactaptin 2 (RL2) has been shown pen- IGOR KIRILYUK1, etrating both cancer and normal cells and inducing apoptosis of cancer cells [1, 2]. Since penetration into cells is OLESYA KRUMKACHEVA2,4, known to be an essential step for biological action of any therapeutic molecules the knowledge of RL2 penetration VLADIMIR RICHTER3, pathways is relevant. Cell-penetrating peptides are known for their effective penetration into cells via direct pene-ELENA BAGRYANSKAYA1 tration through cell membrane or endocytosis and for their ability to deliver cargo molecules. 1N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SB RAS, Novosibirsk, We applied confocal microscopy and CW-EPR methods to examine RL2 penetration into human lung A549 cancer Russia cells. RL2 was labeled by a reduction resistant nitroxide [3-((2,5-dioxopyrrolidin-1-yloxy)carbonyl)-2,2,5,5-tetraeth-2Novosibirsk State University, Physics yl-1-oxyl] spin label to perform in-cell EPR measurements. A549 cells incubated with spin labeled RL2 showed the department, Novosibirsk, Russia 3Institute of Chemical Biology and presence of three EPR spectral components related to different spin label mobility and rotation correlation times Fundamental Medicine, SB RAS, Novosibirsk, differing by more than an order of magnitude. The weight ratio of these components was changing with time Russia 4International Tomography Center, SB RAS, during the cells monitoring by CW-EPR. Confocal microscopy results and EPR spectra simulations allowed us to as-Novosibirsk, Russia sign these components to 1) spin labeled RL2 which is sticking on the membrane surface of endosomes and having spin label rotation correlation time 4.78 ns, 2) spin labeled RL2 with mobility similar to buffer solution and rotation correlation time 2.95 × 10-1 ns, 3) spin labeled amino acids formed in cells due to protein digestion with rotation correlation time 4.1 × 10-2 ns. Thus, CW-EPR and the spin label utilized allowed us to follow the kinetics of different forms of spin labeled RL2 during more than 10 hours after cells incubation with minimal spin concentration 10-60 INTRODUCTION μM in cells. The results of the set of experiments will be shown. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] D. Semenov, A. Fomin, E. Kuligina, O. Koval, V. Matveeva, I. Babkina, N. Tikunova & V. Richter, The protein journal 2010, 29(3), 174-180. [2] O. POSTERS Chinak, A. Shernyukov, S. Ovcherenko, E. Sviridov, V. Golyshev, A. Fomin, I. Pyshnaya, E. Kuligina, V. Richter & E. Bagryanskaya, Molecules, 2019 24(16), 2919. ADVERTISERS Acknowledgements: This study was financially supported by the Ministry of Science and Higher Education of the Russian Federation (grant No. AUTHOR INDEX 14.W03.31.0034). IN009 PARALLEL SESSIONS / INVITED SPEAKER / SMALL MOLECULES I 54 MATHIAS NILSSON MULTIVARIATE AND MULTIWAY NMR School of Chemistry, University of Manchester, Oxford Road, Nuclear magnetic resonance (NMR) spectroscopy is the single tool most widely used by chemists for determining Manchester, U. K. the molecular structures of unknown compounds. It is a wonderfully versatile and sensitive tool, but it has one major drawback: it often struggles to analyze mixtures. Many of the most challenging problems are presented to us as mixtures, so a great deal of effort goes into separating the individual components so that they can be identified and characterized. Fortunately, powerful NMR methods for the analysis of intact mixtures are making its way into the standard set of experiments available to the spectroscopists. One of the most potent is diffusion-ordered spectroscopy (DOSY) in which the signals from different components can be separated by their different diffusion properties. The information available from the basic DOSY experiment can be enhanced by advanced, e.g. multivariate, processing and extended by incorporating information from other sources such as relaxation. This lecture will be focussed on using multivariate and multiway statistics to extract information that is otherwise very hard to obtain. Two-dimensional multivariate techniques, such as PCA, can be very powerful for NMR data, but the so-called rotational ambiguity often results in spectra and components that are linear combinations of the pure components. This can, in suitable cases, be overcome by constraining the solution to match known physical phenomena, such as the shape of the signal decay in a DOSY experiment (SCORE [1,2]) or the form of the kinetics for a chemical reaction. When additional dimensions can be added the data is said to be multiway (e.g. three-way, four-way). When the dimensions are mathematically independent, the rotational ambiguity is no longer affecting the INTRODUCTION data and pure components can be extracted directly from the multi-way decomposition. Examples of this includes, SPONSORS classic chemical reactions [3,4], pathogenic fibril formation [5], and beer analysis [6]. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Colbourne, A. A.; Meier, S.; Morris, G. A.; Nilsson, M. Chem. Commun. 2013, 49, 10510; [2] Nilsson, M.; Morris, G. A. Anal. Chem. 2008, 80, 3777; POSTERS [3] Khajeh, M.; Botana, A.; Bernstein, M. A.; Nilsson, M.; Morris, G. A. Anal. Chem. 2010, 82, 2102; [4] Nilsson, M.; Khajeh, M.; Botana, A.; Bernstein, M. A.; ADVERTISERS Morris, G. A. Chem. Commun. 2009, 1252; [5] Jensen, K. S.; Linse, S.; Nilsson, M.; Akke, M.; Malmendal, A. J. Am. Chem. Soc. 2019, 141, 18649; [6] Dal Poggetto, AUTHOR INDEX G.; Castanar, L.; Adams, R. W.; Morris, G. A.; Nilsson, M. J. Am. Chem. Soc. 2019, 141, 5766. IN010 PARALLEL SESSIONS / INVITED SPEAKER / SMALL MOLECULES I 55 FELIX ROTH, DE NOVO STRUCTURE DETERMINATION OF ORGANIC VOLKER SCHMIDTS, COMPOUNDS FROM MULTI-ALIGNMENT DATA SETS JAN RETTIG, CHRISTINA M. THIELE Residual Dipolar Couplings (RDCs) have proven useful tools in organic structure determination for the determina-Clemens-Schöpf Institute for Organic tion of configuration and/or conformation. [1] Previous approaches for obtaining structures from RDCs in organic Chemistry and Biochemistry, compounds mainly rely on fitting data to structural proposals (e.g., the different diastereoisomers) to verify or fal-Technical University of Darmstadt, Darmstadt, Germany sify them. The process becomes intricate or might even fail in determining the structure, if flexibility plays a (significant) role in the unknown compound. The number of structural proposals grows exponentially with increasing flexibility as then configuration and conformation need to be taken into account (all conformers of each respective diastereoisomer). More recent approaches use RDCs as (pseudo)forces in structure optimisation processes to account for this. [2] Here we present a novel and completely different approach for organic structure determination, which allows for directly obtaining vector and dynamics information from measurements of RDCs in several linearly independent media. While based on an approach from biomolecular NMR [3], it was unclear whether such an approach would be feasible at all for organic compounds. The talk will answer the questions whether in cases of unknown conformation and configuration it is possible to discriminate structures, whether it is possible to induce the many different orientations necessary, and whether it is possible to measure enough couplings per compound. [4] INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] B. Böttcher, C.M. Thiele, Encyclopedia of Magnetic Resonance, 2012, DOI: 10.1002/9780470034590.emrstm1194. [2] G. Cornilescu, R.F. Ramos INVITED AND PROMOTED LECTURES Alvarenga, T.P. Wyche, T.S. Bugni, R.R. Gil, C.C. Cornilescu, W.M. Westler, J.L. Markley, C.D. Schwieters, ACS Chem. Biol. , 2017, 12, 2157-2163; S. Immel, M. Köck, M. Reggelin, Chem. Eur. J. , 2018, 24, 13918-13930; P. Tzvetkova, U. Sternberg, T. Gloge, A. Navarro-Vázquez, B. Luy, Chem. Sci. , 2019, 10, 8774-8791. [3] N.-A. POSTERS Lakomek, K.F.A. Walter, C. Farès, O.F. Lange, B.L. de Groot, H. Grubmüller, R. Brüschweiler, A. Munk, S. Becker, J. Meiler, C. Griesinger, J. Biomol. NMR, 2008, ADVERTISERS 41, 139. [4] F. Roth, V. Schmidts, C.M. Thiele, submitted, preprint available: https://doi.org/10.26434/chemrxiv.14636070.v1; F. Roth, V. Schmidts, J. Rettig, C. AUTHOR INDEX M. Thiele, submitted, preprint available: https://doi.org/10.26434/chemrxiv.14636316.v1 IN011 PARALLEL SESSIONS / INVITED SPEAKER / SMALL MOLECULES I 56 JEAN-NICOLAS DUMEZ DEVELOPMENTS IN ULTRAFAST 2D NMR FOR THE ANALYSIS OF Université de Nantes, CNRS, OUT-OF-EQUILIBRIUM MIXTURES CEISAM, Nantes, France NMR spectroscopy is a powerful tool for the analysis of mixtures, variations are a severe obstacle for reaction monitoring. We showed and the complexity of mixtures often calls for the use of multidi-that different spatial encoding schemes are all affected by the pro- mensional experiments. For samples that evolve in time, the du- cess when a longitudinal axis is used for encoding. On the other ration of these experiments can be a limitation. Ultrafast 2D NMR hand, using a transverse axis for spatial encoding, accurate results based on spatial encoding is the fastest approach to collect 2D NMR can be obtained. This makes it possible to collect 2D spectra in a sin-data [1-2]. It has been used for the monitoring of chemical reactions, gle scan for flow rates of up to 2.5 mL/min that are typical for online and the analysis of hyperpolarised substrates. However, the use of monitoring. spatial encoding is challenging for samples that experience flow. Hyperpolarisation methods provide a much needed sensitivity boost This is a limitation for online monitoring, and for samples hyperpo-for NMR spectroscopy, which could notably benefit to the analysis of larised with methods that involve rapid displacements. mixtures by diffusion-ordered spectroscopy (DOSY). For dissolution Over the past few years, we have developed an array of ultrafast 2D dynamic nuclear polarisation (D-DNP), and Signal Amplification By NMR experiments for the analysis of out-of-equilibrium mixtures Reversible Exchange (SABRE) implemented with the sample-shak-that experience different types of flow. We have characterised the ing approach, rapid displacement of the sample just before acqui-effects of flow on the spatial encoding process, and proposed several sition induces significant internal motion.We showed that, for UF methods to improve the experiments’ accuracy in flow conditions DOSY, the effect of flow can be compensated by a double-diffusion [3-5]. Two types of applications are desribed in this presentation. encoding strategy, as well as by transverse encoding. With UF DOSY INTRODUCTION sequences that compensate or avoid flow effects, accurate results can Online monitoring using flow NMR is a growing area with applica- SPONSORS be obtained in a single scan, thus combining sensivity enhancement tions in organic chemical synthesie and process monitoring, that PROGRAM through hyperpolarisation and extra information through diffusion. would benefit from fast 2D NMR methods. Interference between PROGRAM — BY DAY flow and spatial encoding can result in significant variations of Overall, these methods have the potential to increase the informa- TOPICS the peaks’ intensities of UF 2D 1H-1H correlation spectra, and these tion that can be obtained from mixtures that evolve in time. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] L. Frydman, T. Scherf, A. Lupulescu, Proc. Natl. Acad. Sci. U. S. A. 2002, 99, 15858. [2] M.J. Thrippleton, N. M. Loening, J. Keeler, Magn. Reson. Chem. 2003, 41, 441. [3] C. INVITED AND PROMOTED LECTURES Jacquemmoz, F. Giraud, and J.-N. Dumez, Analyst 2020, 145, 478. [4] L. Guduff, D. Kurzbach, C. van Heijenoort, D. Abergel, J.-N. Dumez. Chem. Eur. J. 2017, .23, 16722. [5] L. Guduff, P. Berthault, POSTERS C. Van Heijenoort, J.-N. Dumez, G. Huber, ChemPhysChem 2019, 20, 392. ADVERTISERS Acknowledgements: This work has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant AUTHOR INDEX agreement no. 801774), the Region Pays de la Loire (Connect Talent) PT009 PARALLEL SESSIONS / PROMOTED TALK / SMALL MOLECULES I 57 JIAQI LU1, NOVEL DARK STATE PHOSPHATE ASSEMBLY FORMATION IN MESOPOTAMIA AQUEOUS SOLUTIONS NOWOTARSKI2, JOSHUA STRAUB3, Phosphate-containing species are in constant flux throughout the (and filterable) component. These findings suggest an entropically ALEXEJ JERSCHOW1, phosphorous cycle and are pooled within the cells of all living or- driven association mechanism, with a small population becoming SONG-I HAN2,4 ganisms. Besides, they also play a potental role in quantum neuro- kinetically trapped. We show that assembly can be facilitated by 1New York University, Department of science. An understanding of the phosphate equilibrium with phos- the addition of depletants and modulated by changing counterion Chemistry, NY, USA 2 phate-containing species and structures under varying conditions salt type, with trends found in line with the Hofmeister series. 31P University of California – Santa Barbara, Department of Chemistry, USA allow for the manipulation of biological energy and the formation NMR Diffusion Oriented Spectroscopy revealed an increase in dif-3University of California – Santa Barbara, and dissolution of biological structures. 31P NMR is commonly used fusion coefficients at elevated temperatures of 70 °C, indicative of Department of Physics, USA 4 to characterize the composition, dynamics and structural proper- dehydration. To further explore the equilibrium of this dark popu- University of California – Santa Barbara, Department of Chemical Engineering, ties of biomolecules and lipid interfaces. While performing 31P NMR lation, we performed chemical exchange saturation transfer exper- USA to investigate the native state of phosphate species as a function of iments, which can identify weakly populated states in proteins [1]. temperature, we encountered peculiar line broadening and relax- Our spectra were wider than expected (by a factor of 2-3), indicating ation effects that cannot be explained by typical dynamical process- exchange with a broad spectroscopic signature population. This es of small molecules. We will present results showing that phos- population appears to increase with temperature as shown by a dip phate containing species reversibly assemble into unreported spec- width increase, thereby supporting that phosphates assemble and troscopically “dark” species, whose fractional population increases are in exchange with detectable phosphate species. with temperature. This study presents a surprising discovery that phosphate mole- INTRODUCTION The NMR-based observation is shown to be consistent with the cules ubiquitously present in biological systems possess a tendency SPONSORS formation of soft phosphate assemblies. Dynamic Light Scattering to form soft assemblies largely invisible to NMR. The conditions in- PROGRAM further provides evidence for the formation of micron-sized as- vestigated under which phosphate-containing molecules in water PROGRAM — BY DAY semblies. The formation of spectroscopically dark states appears assemble should be relevant to a variety of processes that use phos- TOPICS largely reversible (by cooling to 20 °C), with a minor irreversible phates as building blocks. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Vallurupalli, Bouvignies, and Kay, J. Am. Chem. Soc. 2021, 134, 8148-8161. AUTHOR INDEX Acknowledgements: (Matthew P.A. Fisher, Matthew Helgeson) PT010 PARALLEL SESSIONS / PROMOTED TALK / SMALL MOLECULES I 58 RITURAJ MISHRA, ULTRAFAST DIFFUSION-BASED UNMIXING OF 1H-NMR ACHILLE MARCHAND, SPECTRA CORENTIN JAQUEMMOZ, JEAN-NICOLAS DUMEZ Utilization of the spatial-parallelization concept in diffusion-ordered NMR spectroscopy (DOSY), which is known for Université de Nantes, CNRS, CEISAM separation-free identification of mixtures and molecules [1] reduces the acquisition-time of the experiment from UMR6230, Nantes, France few minutes to few hundreds of milliseconds. Recent years have witnessed the use of this spatially encoded-DOSY (SPEN-DOSY) method [2] in mixture analysis via univariate processing methods, where each peak is fitted individually to extract its diffusion constant. However, such analysis on overlapping-peaks of complex mixtures restricts the identification/separation of molecules and leads to wrong values of diffusion constants. This work addresses this problem by using Direct Exponential Curve Resolution Algorithm (DECRA) [3]; a fast multivariate processing algorithm, which separates components of a mixture. This is illustrated with an ethanol-butanol mixture and a sucrose-propanol mixture. Applying DECRA to SPEN data required the design of a radio-frequency (rf) pulse that provides a quadratic spacing of the spatially parallelized gradient area. In addition, clean unmixing of components required additional pre-processing step, to account for the effect of chemical shifts during spatial encoding and during acquisition. Once designed, these new tools are straightforward to implement and use. Combining existing SPEN-DOSY pulse sequence with this newly designed rf pulse, 2D data was acquired for model mixtures, with experiment durations of less than 500 ms. All processing steps are then completed in less than 2 s, out of which only 100 ms required for DECRA. Together these tools provide an “ultrafast” unmixing of 1H NMR spectra, which should be useful for the analysis of INTRODUCTION reaction mixtures and hyperpolarized substrates. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] G. Pagès, V. Gilard, R. Martino and M. Malet-Martino, Analyst 2017, 142, 3771. [2] (a) L. Guduff, I. Kuprov, C. Van Heijenoort and J. N. Dumez, TUTORIAL LECTURE Chem. Commun. 2017, 53, 701; (b) Y. Shrot and L. Frydman, J. Magn. Reson. 2008, 195, 226; (c) M. J. Thrippleton, N. M. Loening and J. Keeler, Magn. Reson. Chem. 2003, 41, 441; (d) N. M. Loening, J. Keeler and G. A. Morris, J. Magn. Reson. 2001, 153, 103. [3] (a) B. Antalek, J. Am. Chem. Soc. 2006, 128, 8402; (b) B. INVITED AND PROMOTED LECTURES Antalek and W. Windig , J. Am. Chem. Soc. 1996, 118, 10331. (c) L. Castañar, G. D. Poggetto, A. A. Colbourne, G. A. Morris and M. Nilsson, Magn. Reson. Chem. POSTERS 2018, 56, 546. ADVERTISERS Acknowledgements: We acknowledge the funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and AUTHOR INDEX innovation program (grant agreement no. 801774), the Region Pays de la Loire (Connect Talent). PT011 PARALLEL SESSIONS / PROMOTED TALK / SMALL MOLECULES I 59 MARIE JURAMY1, MONITORING CRYSTALLIZATION PROCESS IN ROMAIN CHÈVRE1, CONFINED POROUS MATERIALS BY DYNAMIC PAOLO CERREIA VIOGLIO1, FABIO ZIARELLI2, NUCLEAR POLARIZATION SOLID-STATE NMR ERIC BESSON1, STÉPHANE GASTALDI1, Establishing mechanistic understanding of crystallization processes at the molecular level is challenging, as it re-STÉPHANE VIEL1,3, quires both the detection of transient solid phases and monitoring the evolution of both liquid and solid phases as PIERRE THUREAU1, a function of time [1]. Here, we demonstrate the application of dynamic nuclear polarization (DNP) enhanced NMR KENNETH D. M. HARRIS4 AND spectroscopy [2,3] to study crystallization under nanoscopic confinement, revealing a viable approach to interro-GIULIA MOLLICA1 gate different stages of crystallization processes [4]. We focus on crystallization of glycine within the nanometric 1 pores (7-8 nm) of a tailored mesoporous SBA-15 silica material with wall-embedded TEMPO radicals. The results Aix Marseille Univ, CNRS, ICR, Marseille, France show that the early stages of crystallization, characterized by the transition from the solution phase to the first 2 Aix Marseille Univ, CNRS, Centrale Marseille, crystalline phase, are straightforwardly observed using this experimental strategy. Importantly, the NMR sensitivity FSCM, Marseille, France 3 enhancement provided by DNP allows the detection of intermediate phases that would not be observable using Institut Universitaire de France, Paris, France 4 School of Chemistry, Cardiff University, Park standard solid-state NMR experiments. Our results also show that the metastable β polymorph of glycine, which has Place, Cardiff, Wales, U. K. only transient existence under bulk crystallization conditions, remains trapped within the pores of the mesoporous SBA-15 silica material for more than 200 days. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: TUTORIAL LECTURE [1] P. C. Vioglio, P. Thureau, M. Juramy, F. Ziarelli, S. Viel, P. A. Williams, C. E. Hughes, K. D. M. Harris, G. Mollica, J. Phys. Chem. Lett. 2019, 10 (7), 1505–1510. [2] D. A. Hall, D. C. Maus, G. J. Gerfen, S. J. Inati, L. R. Becerra, F. W. Dahlquist, R. G. Griffin, Science 1997, 276 (5314), 930–932. [3] Q. Z. Ni, E. Daviso, T. V. Can, E INVITED AND PROMOTED LECTURES Markhasin, S. K. Jawla, T. M. Swager, R. J. Temkin, J. Herzfeld, R. G. Griffin, Acc. Chem. Res. 2013, 46 (9), 1933–1941. [4] M.Juramy, R. Chèvre, P. Cerreia Vioglio, POSTERS F. Ziarelli, E. Besson, S. Gastaldi, S. Viel, P. Thureau,K. D. M. Harris,G. Mollica, J. Am. Chem. Soc. 2021, In press (DOI: 10.1021/jacs.0c12982) ADVERTISERS Acknowledgements: This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and AUTHOR INDEX innovation programme (grant agreement No 758498). IN012 PARALLEL SESSIONS / INVITED SPEAKER / FIELD-CYCLING NMR RELAXOMETRY 60 DANUTA KRUK DYNAMICS OF IONIC SYSTEMS BY MEANS OF NMR RELAXOMETRY University of Warmia and Mazury in Olsztyn, Department of Nuclear Magnetic Resonance (NMR) relaxometry is highly appre- dynamics, but also access the dimensionality and anisotropy of Physics, Olsztyn, Poland; ciated as a method revealing not only the time-scale but also the these dynamical processes (for instance, reveal the dimensionality danuta.kruk@uwm.edu.pl mechanism of molecular and ionic motion. Fast Field Cycling (FFC) of ionic diffusion). technology has revolutionized this area of NMR relaxation studies The development of batteries is a vital goal of material science and enabling them to be performed as a function of the amplitude of technology. There is a growing awareness that further progress in the magnetic field. Such studies are referred to as “NMR relaxom-the development of electrolyte systems heavily relays on a thor- etry” and they bring a new dimension to NMR by probing different oughgoing understanding of the mechanisms of ionic dynamics. At frequencies of molecular motion. The accessible range of magnet-present, confinement of ionic liquids into nano-porous matrices is ic fields spanning from hundreds of nT to 3T (from about 5kHz to the versatile strategy to overcome disadvantages of liquid electro-above 100MHz, referring to 1H resonance frequency) implies that lytes. Nanoconfined ionic liquids constitute composite (hybrid) sys-one can investigate, in a single experiment, motional processes tems with dynamical properties strongly affected by the nature of across a huge range of time scales from ms to ns. the confinement (solid or polymeric matrices), the size and geom- According to spin relaxation theories, relaxation rates are given etry of the pores and the interactions between ions and pore walls. as linear combinations of spectral density functions being Fourier The price for confining ionic liquids is a significant slowing down transforms of corresponding time correlation functions character-of their mobility, especially long-range translational diffusion de- ising the dynamical processes leading to stochastic fluctuations of termining the conductivity of electrolytes. Consequently, the effort the spin interaction that causes the relaxation. The mathematical in the area of electrolytes based on ionic liquids is strongly focused INTRODUCTION forms of the correlation functions (and, hence, the spectral densi- on developing composites in which fast ionic diffusion is preserved. SPONSORS ties) depend on the mechanism of motion. Consequently, the shape In this lecture an overview of applications of NMR relaxometry to PROGRAM of frequency dependences of spin-lattice relaxation rates (often re- reveal dynamical properties ionic liquid based systems with focus PROGRAM — BY DAY ferred to as relaxation dispersion profiles) is a fingerprint of the na- on the theoretical challenges and achievements will be presented. TOPICS ture of the molecular and ionic motion. In this way one can not only PRIZE LECTURES unambiguously distinguish between translational and rotational PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX Acknowledgements: The work has been supported by National Science Centre, Poland, grant number: 2017/25/B/ST5/02348. IN013 PARALLEL SESSIONS / INVITED SPEAKER / FIELD-CYCLING NMR RELAXOMETRY 61 DAVID J. LURIE1, FAST FIELD-CYCLING MAGNETIC RESONANCE IMAGING LIONEL M. BROCHE1, GARETH R. DAVIES1, Most contrast in conventional MRI arises from differences in T be- We have demonstrated that FFC relaxometry can detect the for- 1 MARY JOAN MACLEOD1, tween normal and diseased tissues. Studies on small tissue samples mation of cross-linked fibrin protein from fibrinogen in vitro [6]. P. JAMES ROSS1, have shown that extra information could be obtained from T -dis- We have also shown that FFC can detect changes in human carti- 1 ROBERT STORMONT1,2 persion measurements (plots of T versus magnetic field), but this in- lage induced by osteoarthritis [7] and differences between normal 1 1 formation is invisible to standard MRI scanners, which operate only tissue and tumour [8]. We have performed in vivo FFC-MRI studies University of Aberdeen, Foresterhill, Aberdeen, United Kingdom at fixed magnetic field ( e.g. 1.5 T). We have developed Fast Field-Cy- on patients with acute ischaemic stroke; FFC-MRI images exhibit- 2GE Healthcare, Waukesha, WI, USA cling (FFC) MRI to exploit T -dispersion as a novel biomarker [1]. ed increased intensity in stroke-affected regions, with maximum 1 contrast typically at the lowest evolution field used (0.2 mT) [9]. FFC relaxometry is conventionally used to measure T -dispersion, 1 All human studies were conducted following approval of the rele- by switching the magnetic field rapidly between levels [2] (polarisa- vant Research Ethics Committees and with the informed consent of tion at high field, followed by evolution (relaxation) at low field, and patients. finally detection at high field. FFC-MRI obtains spatially-resolved T -dispersion data, by collecting MR images at a range of evolution Other work has focused on improving pulse sequences and data 1 magnetic fields [3]. analysis, as well as speeding up the collection of FFC-MRI images [10,11]. Work to improve the hardware and software is ongoing, in- We have built two whole-body human sized FFC scanners, operating cluding the implementation of improved RF coils and receiver coil at detection fields of 0.06 T [4] and 0.2 T [5]. The 0.06 T device uses a arrays [12]. double magnet, with field-cycling being accomplished by switching on and off a resistive magnet inside the bore of a permanent mag- FFC-MRI has significant potential for the generation and use of nov- INTRODUCTION net; this has the benefit of inherently high field stability during the el biomarkers arising from ultra-low field MRI contrast and from SPONSORS detection period. The 0.2 T FFC-MRI system uses a single resistive low- and ultra-low field T -dispersion phenomena. 1 PROGRAM magnet, bringing the advantage of increased flexibility in pulse se- PROGRAM — BY DAY quence programming, at the expense of lower field stability during TOPICS the detection period, necessitating more complex instrumentation. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] D.J. Lurie et al., C. R. Physique 2010, 11, 136-148. [2] R. Kimmich, E. Anoardo, Prog. Nucl. Magn. Reson. Spectrosc. 2004, 44, 257-320. [3] D.J. Lurie, P.J. Ross, L.M. Broche, in: New Developments in NMR No. 18, Kimmich R., ed., Roy. Soc. Chem., UK, 2018, pp 358-384. [4] D.J. Lurie et al., Phys. Med. Biol. 1998, 43, 1877-1886. [5] L.M. Broche et al., Sci. Rep. 2019, 9:10402. [6] INVITED AND PROMOTED LECTURES L.M. Broche et al., Magn. Reson. Med. 2012, 67, 1453-1457. [7] L.M. Broche, G.P. Ashcroft, D.J. Lurie, Magn. Reson. Med. 2012, 68, 358-362. [8] E. Masiewicz et al., Sci. Rep. 2020, 10:14207. [9] POSTERS L.M. Broche et al., 11th Conference on FFC NMR Relaxometry, Pisa, Italy, 2019, p6. [10] P.J. Ross, L.M. Broche, D.J. Lurie, Magn. Reson. Med. 2015, 73, 1120-1124. [11] L.M. Broche, P.J. Ross, K.J. Pine, ADVERTISERS D.J. Lurie, J. Magn. Reson. 2014, 238, 44-51. [12] G.R. Davies et al., ISMRM 27th Annual Meeting, Montreal, Canada, 2019, p1570. AUTHOR INDEX Acknowledgements: This project received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 668119 (project “IDentIFY”). PT012 PARALLEL SESSIONS / PROMOTED TALK / FIELD-CYCLING NMR RELAXOMETRY 62 SVEN BODENSTEDT1, NMR DETECTION AT ULTRALOW FIELD: TAME OR STILL WILD? FRASER HILL-CASEY1, MORGAN MITCHELL1,2, Recently, many groups have jumped onto the bandwagon of ul- Fast-field cycling relaxometry: We recently showed NMR relaxation MICHAEL TAYLER1 tralow-field magnetic resonance following advances in heteronu- measurements [3] in the microtesla field range via a simple setup 1 clear hyperpolarization via para-enriched H and sensitive magne- comprising a water-cooled prepolarizing coil (20 mT), a solenoid (0- ICFO – Institut de Ciencies 2 Fotoniques, The Barcelona Institute tometers (e.g. optically pumped alkali vapors) to detect spin coher- 100 μT, homogeneity ~1 ppk) and an optical magnetometer (<20 fT/ of Science and Technology, ences in the audio band. rtHz sensitivity across 10-1000 Hz frequency range). Coil switching Castelldefels (Barcelona), Spain 2 times below 1 ms allow probing of 1H T and T relaxation rates up ICREA – Institució Catalana The current popularity of ultralow field suggests there is undoubt- 1 2 de Recerca I Estudie Avancats, to several tens of s–1. Examples include study of relaxation in liquid edly “room at the bottom”. However, what remaining problems are Barcelona, Spain molecules near to paramagnetic pore surfaces, and fluids inside really unique to NMR detection in ultralow field, and cannot be thick-walled metal tubing. solved by mainstream approaches, say rf-detection of NMR in high magnetic field? In this presentation we discuss methodological ad- Single scan relaxometry: The NMR line width in ultralow field is vances in the following areas: limited by natural coherence lifetime. We show this property can be used to investigate relaxation of long-lived multinuclear coherenc- Heteronuclear dynamical decoupling at ultralow field: 1H 2H and es for small molecules such as 13CH OH in the strong coupling re-14N 15N isotopic replacement is often performed to mitigate intra- 3 gime between 0-100 μT [3]. We also suggest windowed acquisition molecular dipole-dipole relaxation pathways. In ultralow field, the methods to measure T in the direct dimension. advantage may be outweighed by scalar relaxation effects, poten- 1 tially compromising spectral resolution [1] and polarization trans- fer around level anticrossings [2]. Here we demonstrate trains of dc INTRODUCTION (not ac!) field pulses to induce a spin selectivity that averages out J SPONSORS couplings between 2H and 1H. Composite pulses and a careful choice PROGRAM of phase cycle ensure robustness up to several thousand pulses. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] M.C.D Tayler, L.F. Gladden, J. Magn. Reson. 2019, 298, 101-106. [2] D. Barskiy et al, ChemPhysChem 2017, 18, 1493-1498. [3] S. Bodenstedt, M.W. Mitchell, M.C.D. Tayler, 2020, POSTERS arXiv 2012.05546 ADVERTISERS Acknowledgements: EU H2020 ITN “ZULF” 766402; FEDER Catalunya “QuantumCat” 001-P-001644; la Caixa Junior Leader program LCF/BQ/PI19/116900211; Fundació Privada Cellex; AUTHOR INDEX Fundació Mir Puig. PT013 PARALLEL SESSIONS / PROMOTED TALK / FIELD-CYCLING NMR RELAXOMETRY 63 RUI CORDEIRO1, MARIA J. BEIRA1, TUNING THE 1H NMR PARAMAGNETIC RELAXATION CARLOS CRUZ1, JOÃO L. FIGUEIRINHAS1, ENHANCEMENT AND LOCAL ORDER OF [ALIQUAT]+- MARTA C. CORVO2, PEDRO L. ALMEIDA2,3, ANDREIA A. ROSATELLA4, BASED SYSTEMS MIXED WITH DMSO CARLOS A. M. AFONSO4, CARLA I. DANIEL5, PEDRO J. SEBASTIÃO1 Ionic Liquids are an ever-growing field, not only due to the diversity of known structures, but mainly because, by 1 choosing the proper cation-anion combination, it is possible to tune the physicochemical properties of these sys-Center of Physics and Engineering of Advanced Materials, Departamento de Física, Instituto Superior Técnico, Universidade tems for a wide range of applications. To enable a controlled optimization of these properties, it is important to de Lisboa, Lisbon, Portugal assess both the macroscopic properties of ILs, such as viscosity, as well as to understand these compounds at a 2CENIMAT-Faculdade de Ciências e Tecnologia, Universidade molecular level. Nova de Lisboa, Campus da Caparica, Caparica, Portugal 3Physics Department, ISEL, Lisboa, Portugal 4 More recently, a novel group of ILs, having a paramagnetic metal-based ion incorporated into their structure, was Research Institute for Medicines, Faculty of Pharmacy, Universidade de Lisboa, Lisboa, Portugal described as Magnetic Ionic Liquids (MILs). Some properties of MILs, such as viscosity and diffusion coefficient, can 5LAQV/Requimte, Departamento de Química, Faculdade de be magnetic field dependent, opening a new range of important applications for this type of materials. Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal The presence of a paramagnetic metal in MILs induces an effective NMR relaxation pathway to 1H nuclei, even for metal concentrations in the millimolar range. This effect is the basis for applications such as MRI contrast agents and is known as paramagnetic relaxation enhancement (PRE). This study relies on 1H NMR relaxometry and diffusometry and complementarily uses wide-angle X-rays scattering profiles, which elucidate the effect of DMSO on the local organization of the [Aliquat]+-based ionic liquids. As far INTRODUCTION as the authors know, there are no similar studies performed on these systems, and this is the first ever attempt to SPONSORS access local order dynamics on an ionic liquid [1]. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Cordeiro, R.; Beira, M. J.; Cruz, C.; Figueirinhas, J. L.; Corvo, M. C.; Almeida, P. L.; Rosatella, A. A.; Afonso, C. A. M.; Daniel, C. I.; Sebastião, P. J. INVITED AND PROMOTED LECTURES Int. J. Mol. Sci. 2021, 22 (2), 706. POSTERS Acknowledgements: The NMR spectrometers at IST and FCT NOVA are part of the National NMR network (PTNMR), supported by FCT (ROTEIRO/0031/2013- ADVERTISERS PINFRA/22161/2016). We also acknowledge FCT funding of project UID/DTP/04138/2020 and M.J.B.’s PhD scholarship PD/BD/142858/2018. The authors AUTHOR INDEX would further like to thank European COST Action EURELAX CA15029 (2016–2020). PT014 PARALLEL SESSIONS / PROMOTED TALK / FIELD-CYCLING NMR RELAXOMETRY 64 MARIA ROSARIA RELAXOMETRY OF CANCER: ROLE OF INTRACELLULAR RUGGIERO, WATER LIFETIME AS A TUMOUR BIOMARKER BY IN SIMONA BARONI, VALERIA BITONTO, VIVO FAST FIELD CYCLING SILVIO AIME, SIMONETTA Conventional diagnostic magnetic resonance imaging (MRI) techniques have focused on the improvement of the GENINATTI CRICH spatial resolution by using high magnetic fields (1-7T). High field allows the visualization of small tumour mass but lacks to give a precise evaluation of tumour grading and metastatic potential. Recently, we showed that the intracel-University of Torino, Dep. of Molecular Biotechnology and lular water lifetime represents a hallmark of tumour tissue cells status that can be easily monitored by measuring Health Sciences, Torino, Italy T1 at different and relatively low magnetic field strengths, ranging from 0.2 to 200 mT. A fast exchange through cell membranes indicates a high metabolic rate and thus a high activity of the tumor cells. Thus it is possible to measure the high metabolic pressure by an enhance water exchange with the exterior of the cell. Therefore, intracellular water lifetime can be considered an important tumour biomarker directly depending on the rate of cell proliferation, cell migration and in responding to external stimuli as hypoxia or extracellular acidosis.[1-3] Moreover, currently tumour responses to therapy are monitored primarily by imaging evaluating essentially the decrease of tumor size. This approach, however, lacks sensitivity and can only give a delayed indication of a positive response to treatment. In our study, we propose the use of FFC-NMR to provide relevant information about response to treatment by mon- itoring changes of water exchange rates through cell membranes that are directly dependent on the metabolism alterations caused by the chemo- or radio-therapy. Finally, T relaxation, measured at vey low magnetic fields, can 1 be exploited to support the surgeon in real time margin assessment during breast concerving surgery (BSC). It was INTRODUCTION found that a good accuracy in margin assessment, i.e. a sensitivity of 92% and a specificity of 85%, can be achieved SPONSORS by using two parameters, namely i) the slope of the line joining the R values measured at 0.02 and 1 MHz and ii) the 1 PROGRAM sum of the R values measured at 0.39 and 1 MHz. The obtained results suggest that a simplified, low cost, automat-1 PROGRAM — BY DAY ed instrument might compete well with the currently available tools in BCS margins assessment. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] M Ruggiero et al Angew Chem Int Ed Engl, 2018, 57, 7468-7472; [2] Xin Li et al Magn Reson Med. 2019, 82, 411-424. [3] M Ruggiero et al Molecular POSTERS physics 2019, 1, 1-7. ADVERTISERS Acknowledgements: This project has received funding from the European Union Horizon 2020 research and innovation programme under grant agreement AUTHOR INDEX No 668119 (project “IDentIFY”) IN014 PARALLEL SESSIONS / INVITED SPEAKER / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 65 SHIRA HABER1, STRUCTURAL INSIGHT INTO LAYERED ELECTRODE DANIEL JARDÓN-ÁLVAREZ1, INTERPHASES FROM EXO- & ENDOGENOUS DNP ROSY2, ARKA SAHA2, The electrode – electrolyte interface in rechargeable batteries has a central role in the battery’s performance. A MALACHI NOKED2, plethora of degradation processes, including electrolyte decomposition, structural and mechanical transforma- MICHAL LESKES1 tions lead to loss of energy and power densities. These processes can be controlled through deposition of thin pro-1Department of Molecular Chemistry and tective layers at the electrode interface which prevent degradation, while enabling efficient ion transport across the Materials Science, Weizmann Institute of interface. However, rational design of such interphases is limited by the scarcity of analytical tools that can probe Science, Rehovot, Israel 2Department of Chemistry, Bar Ilan few nanometers thick, heterogenous and disordered layers. University, Ramat Gan, Israel In this talk I will describe how Dynamic Nuclear Polarization (DNP) can be used to efficiently probe thin interphases which are otherwise invisible in NMR. Using different DNP polarization sources – exogenous nitroxide biradicals and endogenous Fe(III) dopants, we are able to gain detailed insight into the composition of the interface as well as its 3D structure. We demonstrate this approach on an alkylated Li Si O interface, deposited through molecular x y z layer deposition, that leads to performance enhancement in high energy cathodes. The combination of exo- and endogenous DNP as well as lithium isotope experiments provide unique insight at the atomic-molecular level which is crucial for designing new materials for high energy battery cells. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN015 PARALLEL SESSIONS / INVITED SPEAKER / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 66 DOMINIK KUBICKI,1,2 SURFACE NMR CRYSTALLOGRAPHY OF METAL HALIDE DANIEL PROCHOWICZ,4 PEROVSKITES JULIA WIKTOR,3* CLARE GREY,2* Surface modification strategies play an essential role in enhancing the optoelectronic performance and stability of SAM STRANKS1* metal halide perovskites.[1] Among the various surface modifiers reported during the past few years, water vapour 1Cavendish Laboratory, Department is one likely to be encountered accidentally. However, it has been challenging to study the formation and elucidate of Physics, University of Cambridge, the atomic-level structure of the surface-bound species, including hydration layers, owing to their extreme dilution. Cambridge, UK 2Department of Chemistry, University The conventional approach employing DNP SENS to study surfaces has proven comparatively inefficient in this of Cambridge, Cambridge, UK class of materials so far.[2] 3Department of Physics, Chalmers University of Technology, To overcome this challenge, we have identified Na+ as a spy cation which binds specifically with the hydrated sur-Gothenburg, Sweden 4Institute of Physical Chemistry, face layer of the perovskite while it has no propensity to interact with the pristine non-hydrated bulk. Owing to its Polish Academy of Sciences, Warsaw, high NMR receptivity (545 times that of 13C), 23Na is perfectly suited for studying dilute surface species, including Poland real-time monitoring of hydration. We employ a combination of 23Na MAS NMR and chemical shift calculations to elucidate the speciation of sodium within the hydrated surface layer and show that it provides a unique atomic-level picture of its complex interior. This strategy broadens the range of materials research questions that have been successfully addressed using sol-id-state MAS NMR in the context of metal halide perovskite research.[3,4] INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Chen, B., N. Rudd, P., Yang, S., Yuan, Y. & Huang, J. I, Chem. Soc. Rev. 2019, 48, 3842–3867. [2] Hanrahan, M. P., Men, L., Rosales, B. A., Vela, J. & Rossini, A. J., Chem. Mater. 2018, 30, 7005–7015. [3] Piveteau, L., Morad, V. & Kovalenko, M. V., J. Am. Chem. Soc. 2020. [4] Kubicki, D. J., Stranks, S. D., Grey, C. INVITED AND PROMOTED LECTURES P. & Emsley, L. NMR Nat. Rev. Chem. 2021 in press. POSTERS Acknowledgements: This work has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie ADVERTISERS Skłodowska-Curie grant agreement No. 841136. S.S. acknowledges the Royal Society and Tata Group (UF150033). The work has received funding from the AUTHOR INDEX European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (HYPERION, grant agreement No. 756962). PT015 PARALLEL SESSIONS / PROMOTED TALK / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 67 JENNIFER GÓMEZ1, OBSERVATION OF QUADRUPOLAR NUCLEI IN SOLIDS BY HIROKI NAGASHIMA2, CONTROLING INTERNUCLEAR TRANSFERS NGHIA TUAN DUONG1,3, MINGJI ZHENG4, Half-integer quadrupolar nuclei, such as 11B, 17O and 27Al, represent were combined with DNP to detect the NMR signals of quadrupo-SHAOHUI XIN4, about two thirds of stable NMR-active nuclei and are present in or- lar isotopes with low natural abundance (17O) and low gyromagnet- QIANG WANG4, ganic, inorganic and biological solids . Nevertheless, the NMR ob- ic ratio (47,49Ti, 67Zn, 95Mo), notably near the surface of the materials. JULIEN TRÉBOSC5, servation of these nuclei in solids remains often more challenging Furthermore, we acquired high-resolution NMR spectra of surface FENG DENG4, than those of spin-1/2 nuclei. This difficulty comes from the larger 17O nuclei by combing these transfers with multiple-quantum MAS GAËL DE PAËPE3, size of the density matrix for quadrupolar nuclei as well as the large sequence. JEAN-PAUL AMOUREUX1,6, anisotropic quadrupolar interaction, which complicates the spin OLIVIER LAFON1,7 For the observation of homonuclear proximities between quadru- dynamics. In particular, the cross-polarization under magic-an- 1 polar nuclei, we have shown that the high robustness of symme- Univ. Lille, CNRS, UCCS, Lille, France. gle spinning (CPMAS) lacks of robustness for quadrupolar nuclei, 2AIST, Tsukuba, Japan, try-based recoupling stems from the role of offset for the 3 which limits the observation of proximities with other isotopes but Univ. Grenoble Alpes, CEA, IRIG-MEM, reintroduction of dipolar interactions [4]. Furthermore, for this re- Grenoble, France. also the enhancement of NMR signals by dynamic nuclear polariza- 4 coupling, the intensity of correlations is reduced on-resonance by Chinese Academy of Sciences, WIPM, China. tion (DNP). 5Univ. Lille, CNRS, IMEC, Lille, France. the creation of unwanted coherences. We also investigated pulse se- 6Bruker Biospin, France. In the past year, we made significant progresses to circumvent these quences for the observation of heteronuclear proximities between 7IUF, France. issues. We introduced robust symmetry-based heteronuclear di- half-integer quadrupolar nuclei [5]. We notably showed that the polar recoupling built from adiabatic or composite inversion puls- recoupling scheme must be applied to the channel for which the es to transfer the polarization of protons to quadrupolar nuclei at magnetization is parallel to the B field. 0 INTRODUCTION MAS frequencies ranging from 10 to 60 kHz [1-3]. These transfers SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] H. Nagashima, J. Trébosc, Y. Kon, K. Sato, O. Lafon, J.-P. Amoureux, J. Am. Chem. Soc., 2020, 142, 10659. [2] H. Nagashima, J. Trébosc, Y. Kon, O. Lafon, J.-P. Amoureux, Magn. POSTERS Reson. Chem., 2021, in press doi:10.1002/mrc.5121. [3] J. S. Gómez, A. G.M. Rankin, J. Trébosc, F. Pourpoint , Y. Tsutsumi, H. Nagashima, O. Lafon, J.-P. Amoureux Magn. Reson. submitted doi: ADVERTISERS 10.5194/mr-2021-29. [4] N. T. Duong, D. Lee, F. Mentink-Vigier, O. Lafon, G. De Paëpe Magn. Reson. Chem., 2021, in press doi: 10.1002/mrc.5142 [5] M. Zheng, S. Xin, Q. Wang, J. Tébosc, J. Xu, G. AUTHOR INDEX Qi, N. Feng, O. Lafon, F. Deng Magn. Reson. Chem., 2021, in press doi: 10.1002/mrc.5163 PT016 PARALLEL SESSIONS / PROMOTED TALK / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 68 TAMAR WOLF, QUADRUPOLAR ISOTOPE CORRELATION SPECTROSCOPY: MICHAEL J. JAROSZEWICZ, RESOLVING OVERLAPPING WIDELINE NMR SPECTRA LUCIO FRYDMAN Department of Chemical and Biological OF QUADRUPOLAR NUCLEI UNDER STATIC CONDITIONS Physics, Weizmann Institute of Science, Rehovot, Israel Solid-state NMR spectra of quadrupolar nuclei provide a wealth of two quadrupolar isotopes of the same element. Several elements information pertaining to molecular structure, dynamics, chemical that are important and ubiquitous across materials science, phar- identity, and purity. Extracting this information, which is encoded maceuticals, and inorganic chemistry possess at least two NMR-ac-in the quadrupolar coupling constant ( C ), the asymmetry param- tive quadrupolar isotopes, which are amenable to QUICSY. Exam- Q eter (η ) and the chemical shift parameters, is generally straight- ples include the isotopes of chlorine (35,37Cl) and bromine (79,81Br), Q forward when investigating samples that possess a single chemical which are often present in active pharmaceutical ingredients [4]; site. However, significant difficulty in spectral analysis arises when the isotopes of gallium (69,71Ga), which comprise semiconductors investigating samples possessing multiple non-equivalent chemi-and metal-organic frameworks [5], and the isotopes of rubidium cal sites, which give rise to overlapping NMR spectra of each site, (85,87Rb). This approach is demonstrated here using two model rubid-and drastically reduce individual site resolution. Current methods ium-containing compounds, RbClO and Rb SO , which possess one 4 2 4 that address these problems are useful only in cases where the C s and two chemical sites, respectively. The resulting 2D QUICSY NMR Q are relatively small, under magic-angle spinning (MAS) [1-2], yet are spectra, provide rich information about the magnitude and geome- not well suited for large C s –for which static acquisition ( i.e., no try of the quadrupolar and the chemical shift interactions, as well as Q MAS) is preferable [3]. Hence, there is a necessity for robust meth- details concerning the relative orientation of their respective inter- ods capable of resolving and deconvolving overlapped broad NMR action tensors. We validate our results by comparing experimental INTRODUCTION spectra that are characterized by large C values. 2D QUICSY spectra to 2D analytical calculations based on previous Q SPONSORS literature parameters. Thus, 2D QUICY NMR might offer a way to ex- PROGRAM To this end, herein we introduce QUadrupolar Isotope Correlation tract the NMR parameters for overlapping wideline spectra of quad- PROGRAM — BY DAY SpectroscopY (QUICSY) as a potential method for resolving over- rupolar nuclei when conventional methods cannot be applied. TOPICS lapped NMR spectra under static conditions, by correlating – via PRIZE LECTURES cross polarization – the central-transition NMR spectra arising from PLENARY LECTURES TUTORIAL LECTURE References: [1] A. Medek, J.S. Harwood, L. Frydman, J.Am.Chem.Soc. 1995, 117, 12779-12787. [2] Z. Gan, J.Am.Chem.Soc. 2000, 122, 3242-3243. [3] M.J. Jaroszewicz, L. Frydman, R. Schurko, J.Phys. INVITED AND PROMOTED LECTURES Chem.A. 2017, 121, 51-65. [4] M. Hildebrand et al. , CrystEngComm. 2014, 16, 7334-7356. [5] R. Hajjar et al. , Chem. Mater. 2011, 23, 39-47. POSTERS Acknowledgements: We are grateful to Prof. Shimon Vega for important discussions regarding this project. This work was funded by the Israel Science Foundation grant 965/18, by the ADVERTISERS Minerva Foundation (Germany), and the generosity of the Perlman Family Foundation. T.W. acknowledges the Clore Foundation for a graduate student fellowship. L.F. holds the Bertha and AUTHOR INDEX Isadore Gudelsky Professorial Chair and Heads the Clore Institute for High-Field Magnetic Resonance Imaging and Spectroscopy-whose support is also acknowledged. PT017 PARALLEL SESSIONS / PROMOTED TALK / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 69 DOROTHEA WISSER1, ADSORPTION OF PHOSPHATE ON γ-ALUMINA ADRIAN HÜHN2, ELUCIDATED BY DNP-ENHANCED SOLID-STATE NMR MANUEL CORRAL VALERO3, TEDDY ROY3, SPECTROSCOPY AND DFT CALCULATIONS MICKAËL RIVALLAN3, LEONOR DUARTE3, Transitional γ-aluminas are widely used in industrial processes as heterogeneous catalysts and catalyst supports. ANNE LESAGE4, Numerous empirical studies have shown that inorganic additives, such as phosphate, have a strong impact on the CARINE MICHEL2, formation of catalytically active phases on γ-alumina and on the resulting activity of the material. [1-3] PASCAL RAYBAUD2,3 However, an atomic-scale description of the chemical interactions of phosphates with γ-alumina is still missing, 1Erlangen Center for Interface Research and but is essential to obtain a rational understanding of the role of this additive. The combination of MAS NMR and Catalysis, Friedrich-Alexander-Universität DFT calculations has recently allowed us to disentangle the nature and location of surface hydroxyl groups on fac-Erlangen-Nürnberg, Erlangen, 2Univ Lyon, ENS de Lyon, CNRS UMR ets and edges of γ-alumina crystallites.[4] Here, we present an unprecedented insight into phosphate speciations 5182, Université Claude Bernard Lyon 1, and localizations on γ-alumina by combining cutting-edge Dynamic Nuclear Polarization Surface Enhanced NMR Laboratoire de Chimie, Lyon, France, 3 Spectroscopy (DNP SENS) techniques with Density Functional Theory (DFT) calculations. Using 31P double- and tri-IFP Énergies nouvelles, Solaize, France, 4Centre de RMN à Très Hauts Champs, ple-quantum filtered NMR experiments, the presence of bi- and polyphosphates is investigated. By 27Al-31P dipolar-Université de Lyon, Villeurbanne, France. and scalar-based correlation spectra, Al-O-P connectivities are established. These experimental results are combined with DFT calculations, which allow to determine in detail the thermodynamically most stable binding modes of phosphate and preferential binding sites on different crystallite facets. 31P NMR chemical shifts were calculated and compared to the experimental data. The evolution of phosphate speciations and binding sites is studied in a INTRODUCTION broad range of phosphate coverages up to 4.6 P/nm2. The results will support future knowledge-driven develop- SPONSORS ment of catalytic materials, by better targeting surface modifier concentration and binding modes in view of their PROGRAM specific role in the catalysis itself or in nanoparticle formation. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] K. Bourikas, C. Kordulis, A. Lycourghiotis, Catal. Rev. 2006, 48, 363–444. [2] P. Munnik, P.E. de Jongh, K.P. de Jong, Chem. Rev. 2015, 115, 6687– ADVERTISERS 6718. [3] M. Corral Valero, P. Raybaud, J. Catal. 2020, 391, 539–547. [4] A. T. Batista, D. Wisser, T. Pigeon, D. Gajan, F. Diehl, M. Rivallan, L. Catita, A.-S. Gay, A. AUTHOR INDEX Lesage, C. Chizallet, P. Raybaud, J. Catal. 2019, 378, 140–143. IN016 PARALLEL SESSIONS / INVITED SPEAKER / HYPERPOLARIZATION 70 BJÖRN CORZILIUS LOCALIZED DNP ENHANCEMENT IN BIOMOLECULES Institute of Chemistry and Department of Life, Light & With increasing complexity of biomolecular assembly, sensitivi- experimental evidence for this elusive DNP distance dependence Matter, University of Rostock, ty as well as specificity play a major role in NMR-based structural on a ubiquitin model protein spin-labeled with a Gd3+ chelate tag Rostock, Germany biology. Dynamic nuclear polarization (DNP) has shown tremen- [3]. By correlating the 15N build-up rates with computationally as- dous potential to increase sensitivity in numerous applications [1]. sisted structural modeling of the paramagnetically labeled protein, Even though in conventional DNP experiments uniform signal en-the theoretically r−6 dependence is observed. However, the para- hancements are typically obtained, DNP itself can act as a source of magnetic Gd3+ generates a bleaching volume with a radius of 12 Å specificity as well [2]. In this presentation, two methods allowing to within which no nuclear spin contributes to the measured rate. This introduce a large degree of specificity to DNP-enhanced MAS NMR opens up the possibility to extract distance restraints directly from spectra will be presented. direct DNP which can be further used in structural modeling. The first method utilizes the distance dependence of the dipolar hy- The second method employs efficient indirect DNP (via 1H), but perfine interaction between the electron spin (source) and nuclear introduces methyl groups as localized polarization transfer path-spin (target). The hyperfine interaction is mediating the initial step ways between 1H and 13C. The introduction of specifically 13C-labeled of the complex mechanism of the overall DNP transfer. By micro-methyl groups via different biomolecular methods enables the dis- wave irradiation, electron-nuclear coherences are generated which crete placement of antennas for hyperpolarization within the car-finally result in nuclear hyperpolarization. If subsequent spin dif- bon network. Based on heteronuclear cross-relaxation, SCREAM- fusion is restricted, this transfer dynamic can act as a measure for DNP (specific cross-relaxation enhancement by active motions un-hyperfine interaction. However, the competing paramagnetically der DNP) can thus probe the local environment around these highly INTRODUCTION enhanced spin-lattice relaxation counteracts the creation of a po- dynamic functions [4]. This is especially interesting in the context SPONSORS larization gradient, theoretically leading to uniform, distance-in- of biomolecular complexes and specific contacts between binding PROGRAM dependent DNP enhancement. Nevertheless, the direct-DNP partners [5]. Examples and applications on proteins, RNA, and ri- PROGRAM — BY DAY build-up rate can act as a direct measure of this interaction and can bonucleoproteins (RNPs) will be presented and future implications TOPICS thus yield distance information in biomolecules. We will present discussed. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] A. S. Lilly Thankamony, J. J. Wittmann, M. Kaushik and B. Corzilius, Prog. Nucl. Magn. Reson. Spectrosc. 2017, 102-103, 120. [2] R. Rogawski and A. E. McDermott, Arch. Biochem. INVITED AND PROMOTED LECTURES Biophys. 2017, 628, 102. [3] J. Heiliger, T. Matzel, E. C. Çetiner, H. Schwalbe, G. Kuenze and B. Corzilius, Phys. Chem. Chem. Phys. 2020, 22, 25455. [4] V. Aladin and B. Corzilius, eMagRes 2020, POSTERS 9, 1. [5] V. Aladin, M. Vogel, R. Binder, I. Burghardt, B. Suess and B. Corzilius, Angew. Chem. Int. Ed. 2019, 58, 4863. ADVERTISERS Acknowledgements: Funded through Deutsche Forschungsgemeinschaft (DFG grants CO 802/2-1, CO 802/2-2, CO 802/2-4 and INST 264/177-1). Further financial support from the BMRZ AUTHOR INDEX (Frankfurt) is acknowledged. IN017 PARALLEL SESSIONS / INVITED SPEAKER / HYPERPOLARIZATION 71 MEGHAN E. HALSE, ENHANCING PORTABLE NMR SPECTROSCOPY USING SEAN CAWKWELL, PARAHYDROGEN HYPERPOLARISATION SIMON DUCKETT, FRASER HILL-CASEY, Increasing NMR sensitivity through hyperpolarisation holds great promise for integration with portable NMR spec-JAMES MCCALL, trometers, whose relatively low magnetic fields (≤ 2 T) limit both signal strength and chemical shift dispersion. ALASTAIR ROBINSON, Para hydrogen-induced polarisation (PHIP) methods are of particular interest because they can deliver high levels of MATHEUS ROSSETTO, hyperpolarisation (up to tens of %), which are independent of the detection field and build up over a period of sec-AMINATA SAKHO onds. Importantly, PHIP methods can be implemented with relatively inexpensive and compact instrumentation. Department of Chemistry, University The key to unlocking the hyperpolarisation potential of para hydrogen ( p-H ) is a chemical reaction that breaks the 2 of York, York, UK symmetry of the protons in p-H that exist in a singlet state, either through hydrogenation of an unsaturated mole-2 cule (e.g. an alkene or alkyne) [1] or through reversible binding to a metal centre (the signal amplifcation by reversible exchange (SABRE) approach) [2]. Both hydrogenative PHIP and non-hydrogenative SABRE have the potential to unlock new applications for low-field, portable NMR spectroscopy. In addition, ultra-low-field NMR can provide insights into the spin dynamics of p-H hyperpolarisation and so can be used to develop new optimised polarisation 2 transfer strategies. This presentation will explore a range of applications for parahydrogen-enhanced portable NMR including: mon- itoring photochemical reactivity using benchtop NMR detection and p-H hyperpolarisation, observing the spin 2 dynamics of a range of iridium di-hydride complexes that provide continuous hyperpolarisation in the ultra-low- field regime through reversible exchange with p-H , and the use of field-cycled NMR with Earth’s field detection [3] 2 INTRODUCTION to explore SABRE polarisation transfer mechanisms in strongly-coupled 1H-19F spin systems. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] Eisenschmid et al. J. Am. Chem. Soc. 1987, 109, 8089; [2] Adams R.W. et al. , Science 2009, 323, 1708-11; [3] Hill-Casey et al. , Molecules 2019, ADVERTISERS 24, 4162; AUTHOR INDEX Acknowledgements: UKRI EPSRC (EP/M020983/1, EP/R028745/1); EU Horizon 2020 Marie Skłodowska-Curie grant agreement No. 766402 IN018 PARALLEL SESSIONS / INVITED SPEAKER / HYPERPOLARIZATION 72 DENNIS KURZBACH INTEGRATED DISSOLUTION DNP, NMR, AND EPR University Vienna, Faculty of TO DERIVE STRUCTURE-ACTIVITY RELATIONSHIPS OF Chemistry, Institute of Biological Chemistry, Vienna, Austria INTRINSICALLY DISORDERED PROTEINS Transcription factors (TFs) play a critical role in numerous physio- protein. It forms a stiff elongated coiled-coil helical dimer, adjoin- logical and pathological processes. Many important TFs are charac- ing a disordered N-terminal domain that houses the DNA binding terized by a notably high degree of conformational plasticity, sug- site. The dysregulated expression and activity of MYC and MAX are gesting that intrinsic disorder (ID) may be essential to fulfill their of central importance in several tumor types. However, the charac-functional roles. terization of MAX is complicated, in particular under physiologi- cal conditions and concentrations, by weak signal intensities and Many fold into stable structures only upon DNA binding; in the crowded spectra, owing to both the slow and anisotropic rotation of DNA-free state, conformational heterogeneity often renders char-the dimer and the intrinsic disorder of the DNA-binding site. As a acterization of these substrates experimentally challenging. Hence, consequence, the functional role of intrinsic disorder for MAX has atomic-level details of their structures, dynamics, and interactions remained unclear until now. often lack despite their considerable medicinal importance. Here we address this question by integrating dissolution dynamic nu- Our studies relate in particular to the long-standing dilemma be- clear polarization (D-DNP) [1], carbon direct-detected NMR exper- tween spatial plasticity of transcription factors and their generally iments, paramagnetic relaxation enhancements, MD simulations, high binding affinities, which counteract relocalization in DNA-and EPR nanoscale distance measurements. With this approach, we bound states. ‘Facilitated diffusion’ and ‘facilitated dissociation’ could develop detailed models of the role of intrinsically disordered models try to answer how TFs move along DNA, how they pass other INTRODUCTION TF domains and how their heterogeneous conformational sampling bound molecules while diffusing along the double-strands, and how SPONSORS modulates transcriptional activity [2-4]. they detach from DNA strands. We show how MAX solves these tasks PROGRAM by involving its intrinsically disordered domain enabling (i) rapid PROGRAM — BY DAY We focus on MAX (MYC associated transcription factor X) as a mod- switching between loosely and tightly DNA-bound states and (ii) wid- TOPICS el system, a member of the basic helix-loop-helix leucine zipper ening its sampling space to recruit DNA to the binding site efficiently. PRIZE LECTURES (bHLH-LZ) family, is a well-known example of a partially disordered PLENARY LECTURES TUTORIAL LECTURE References: [1] J. H. Ardenkjaer-Larsen, B. Fridlund, A. Gram, G. Hansson, L. Hansson, M. H. Lerche, R. Servin, M. Thaning and K. Golman, Proc Natl Acad Sci, 2003, 100, 10158-10163. [2] G. INVITED AND PROMOTED LECTURES Sicoli, H. Vezin, K. Ledolter, T. Kress and D. Kurzbach, Nucleic Acids Res. 2019, 47, 5429-5435. [3] G. Sicoli, T. Kress, H. Vezin, K. Ledolter and D. Kurzbach, J Phys Chem Lett, 2020, 8944-8951. POSTERS [4] G. Kizilsavas, K. Ledolter and D. Kurzbach, Biochemistry 2017, 56, 5365-5372. ADVERTISERS Acknowledgments: Based on contributions from G Sicoli, H Vezin, T Kress, LM Epasto, K Che, F Kozak, A Selimović, M Negroni, K Ledolter. Funding: ERC-StG 801936, FWF P33338-N, AWS AUTHOR INDEX Prototype Funding. PT018 PARALLEL SESSIONS / PROMOTED TALK / HYPERPOLARIZATION 73 SVETLANA PYLAEVA1, MIXED-VALENCE POLARIZING AGENTS FOR EFFICIENT ANDREI GURINOV2, MARC OVERHAUSER EFFECT DNP IN INSULATING SOLIDS AT BALDUS2, MICHAEL RÖMELT3, HOSSAM ELGABARTY1, HIGH MAGNETIC FIELDS THOMAS D. KÜHNE1, BENEDIKT SIELAND1, Overhauser effect DNP in insulating solids [1] has a number of beneficial properties for applications: it requires JAN PARADIES1, ANDREI less microwave power than other methods, the enhancement was shown to scale favourably with magnetic field KUZHELEV4, THOMAS strength [2]. PRISNER4, KONSTANTIN L. Mixed-valence character of the BDPA radical was recently investigated using state-of-the-art quantum-chemistry IVANOV5 methods. We have vigorously proven that the radical belongs to a specific class of mixed valence compounds: spin 1University of Paderborn, Department of density is localized on one part of the molecule, but can tunnel to the other side with a frequency in a range of hun-Chemistry, Paderborn, Germany 2 dreds of GHz [3]. Interestingly, same conclusions for BDPA were reached in 1975 by Watanabe et al. [4]. Utrecht University, Bijvoet Center, Utrecht, the Netherlands 3 Now we have performed theoretical scanning of large number of mixed-valence molecules, looking for ones with Ruhr University Bochum, Lehrstuhl für Theoretische Chemie, Bochum, Germany properties similar to those of the BDPA (g-tensor anisotropy, large values of hyperfine coupling constants, similar 4Goethe University Frankfurt am Main, electron transfer rate). Those molecules have been then synthesized and investigated by high field EPR. Their DNP Institute of Physical and Theoretical Chemistry, Frankfurt am Main, Germany field profiles were measured at 18.8 Tesla (527 GHz) at 100 K in tetrachloroethane (90D:10H) matrix under 8kHz 5International Tomography Center, MAS, concentration of the radicals was 10 mM. We have indeed observed large signal enhancement for all three Novosibirsk, Russia radicals when irradiated on EPR resonance. Other properties of the signal, such as MAS and microwave power de- pendence are similar to the BDPA [5]. INTRODUCTION SPONSORS Our finding opens a whole class of mixed-valence radicals to prospective applications as polarizing agents at ul- PROGRAM tra-high magnetic fields. Organic mixed-valence compounds make up a large and well-studied group of molecules, PROGRAM — BY DAY their versatile nature allows for adaptation for various applications. TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] T. V. Can, M. A. Caporini, F. Mentink-Vigier, B. Corzilius, J. J. Walish, M. Rosay, W. E. Maas, M. Baldus, S. Vega, T. M. Swager, R. G. Griffin, J. Chem. TUTORIAL LECTURE Phys. 2014, 141, 064202. [2] M. Lelli, S. R. Chaudhari, D. Gajan, G. Casano, A. J. Rossini, O. Ouari, P. Tordo, A. Lesage, L. Emsley, L. J. Am. Chem. Soc. 2015, 137, 14558. [3] S. Pylaeva, P. Marx, G. Singh, T. D. Kühne, M. Roemelt, H. Elgabarty, J. Phys. Chem. A, 2021, 125, 867. [4] K. Watanabe, J. Yamauchi, H. Ohya- INVITED AND PROMOTED LECTURES Nishiguchi, Y. Deguchi, K. Ishizu, Bull. Inst. Chem. Res., Kyoto Univ. 1975 , 53(2), 161. [5] A. Gurinov, B. Sieland, A. Kuzhelev, H. Elgabarty, T. D. Kuehne, T. Prisner, POSTERS J. Paradies, M. Baldus, K. L. Ivanov, S. Pylaeva, Angew. Chem. Int. Ed. 2021, https://doi.org/10.1002/anie.202103215. ADVERTISERS Acknowledgements: This work has been supported by iNEXT-Discovery, grant number 871037, funded by the Horizon 2020 program of the European AUTHOR INDEX Commission. PT019 PARALLEL SESSIONS / PROMOTED TALK / HYPERPOLARIZATION 74 THÉO EL DARAÏ1,2, POROUS FUNCTIONALIZED POLYMERS SAMUEL F. COUSIN1, ENABLE GENERATING AND TRANSPORTING QUENTIN STERN1, MORGAN CEILLIER1, HYPERPOLARIZATION JAMES KEMPF3, DMITRY ESHCHENKO3, More than fifteen years after the invention of dissolution dynamic nuclear polarization (d-DNP) [1], it was demon-ROBERTO MELZI3, strated that d-DNP could potentially be performed remotely, off-site [2], thus without the need of a polarizer on-site. MARC SCHNELL3, In our group, we have in the past years, merely been working at improving efficiency [3], compatibility [4], and LAURENT GREMILLARD4, repeatability [5] of d-DNP. Our candid objective basically was to enable (or at least to improve) applications; none-AURÉLIEN BORNET1, theless it has led us to develop a new concept i) to dramatically extend hyperpolarization lifetimes from minutes to JONAS MILANI1, days and, ii) to enable transport to far distant MRI or NMR sites [2]. BASILE VUICHOUD1, We are now generalizing this new concept to a broad range of systems, such as neat endogenous tracers, mixtures OLIVIER CALA1, of metabolites, or amino acids, by developing new hyperpolarizing solids such as our silica-based HYPSO materials, DAMIEN MONTARNAL2, or more recently hyperpolarizing porous polymers (HYPOP). These can be impregnated with arbitrary solutions SAMI JANNIN1 that are then hyperpolarized efficiently and stored and transported over hours, before being melted and released. 1CRMN, Villeurbanne, France, 2Univ Lyon, CPE Lyon, Lyon, France, We will present DNP results on the very first generation of these new materials [6]. We’ll also show how the porosity 3Bruker Biospin, and morphology of this material can be tuned to further improve performances. Finally, we also show how hyperpo-4Univ Lyon, INSA Lyon, Villeurbanne, France larization lifetimes can be further extended by treating the HYPOP surfaces before impregnation, and performing cold-crystallization of the sample after impregnation. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES References: [1] Ardenkjaer-Larsen, J. H. et al. Proc. Natl. Acad. Sci. 2003, 100, 10158–10163. [2] Ji, X. et al. Nat. Commun. 2017, 8; Capozzi, A. et al. Nat. Commun. PLENARY LECTURES 2017, 8, 15757. [3] Bornet, A. et al. J. Phys. Chem. Lett. 2013, 4, 111–114 ; Bornet, A. et al. Chem. Phys. Lett. 2014,602, 63–67 ; Bornet, A. et al. Phys. Chem. Chem. TUTORIAL LECTURE Phys. 2016, 18, 30530–30535. [4] Gajan, D. et al. Proc. Natl. Acad. Sci. USA 2014, 111, 14693–14697 ; Cavaillès, M. et al. Angew. Chemie Int. Ed. 2018 ; Vuichoud, B. et al. J. Phys. Chem. B 2014, 118, 1411–1415. [5] Bornet. A. et. al. Anal. Chem. 2016, 88, 6179-6183. [6] El Daraï, T. et. al. https://doi.org/10.21203/rs.3.rs-123790/ INVITED AND PROMOTED LECTURES v1 under review. POSTERS Acknowledgements: This research was supported by ENS-Lyon, the French CNRS, Lyon 1 University, the Institute of Chemistry at Lyon (ICL), the European ADVERTISERS Research Council under the European Union’s Horizon 2020 research and innovation program (ERC Grant Agreements No. 714519 / HP4all and Marie AUTHOR INDEX Skłodowska-Curie Grant Agreement No. 766402 / ZULF). PT020 PARALLEL SESSIONS / PROMOTED TALK / HYPERPOLARIZATION 75 FREDERIC THE DISTANCE BETWEEN G-TENSORS OF NITROXIDE MENTINK-VIGIER1, BIRADICALS GOVERNS MAS-DNP PERFORMANCE IN THIERRY DUBROCA1, JOHAN VAN TOL1, THE BTUREA FAMILY SNORRI TH. SIGURDSSON2 Limited sensitivity of solid-state NMR experiments makes it challenging to study systems where the isotope of in-1 terest is in low concentration. To overcome the limitations, bis-nitroxide radicals [1] are common polarizing agents National High Magnetic Field Laboratory, Florida State (PA) that are used to enhance the nuclear polarization during Magic Angle Spinning Dynamic Nuclear Polariza- University, Tallahassee, FL, USA, tion (MAS-DNP) [2,3]. These biradicals efficiently increase the proton spin polarization through the Cross-Effect 2Science Institute, University of mechanism [4,5]. The relative orientation of the bis-nitroxide moieties is critical to ensure efficient polarization Iceland, Reykjavik, Iceland transfer. [6] Recently, we have defined a new quantity, the distance between g-tensors, that correlates the relative orientation of the nitroxides with the ability to polarize the surrounding nuclei. [7] Here we analyse experimentally and theoretically a series of biradicals belonging to the bTurea family, namely bcTol [8], AMUPol [9] and bcTol-M [10]. They differ by the degree of substitution on the urea bridge that connects the two nitroxides. Using quantitative simulations developed for moderate MAS frequencies, we show that these modifications mostly affect the relative orientations of the nitroxide, i.e. the length and distribution of the distance between the g-tensors, that in turn impacts both the steady state nuclear polarization/depolarization as well as the build-up times. The doubly substituted urea bridge favours a large distance between the g-tensors, which enables bcTol-M to provide ϵ > 200 on/off at 14.1 T/600 MHz/395 GHz with build-up times of 3.8 s using a standard homogenous solution. The methodology described herein was used to show how the conformation of the spirocycles flanking the nitroxides in the recently INTRODUCTION described c- and o-HydrOPol biradicals [11] affects the relative orientation of the nitroxides and thereby the polar- SPONSORS ization performance. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES References: [1] Hu, K.-N., H. Yu, T. M. Swager, et al., J. Am. Chem. Soc. 2021, 126, 10844. [2] Hall, D. A., D. C. Maus, G. J. Gerfen, et al., Science, 1997, 276, 930 PLENARY LECTURES (1997). [3] Rosay, M., L. Tometich, S. Pawsey, et al. , Phys. Chem. Chem. Phys. 2010 12, 5850. [4] Mentink-Vigier, F., U. Akbey, Y. Hovav, et al., J. Magn. Reson. 2012, TUTORIAL LECTURE 224, 13. [5] Thurber, K. R. and R. Tycko, J. Chem. Phys. 2012, 137, 084508. [6] Ysacco, C., E. Rizzato, M.-A. Virolleaud, et al., Phys. Chem. Chem. Phys. 2010, 12, 5841. [7] Mentink-Vigier, F., Phys. Chem. Chem. Phys. 2020, 22, 3643. [8] Jagtap, A. P., M.-A. Geiger, D. Stöppler, et al., Chem. Commun. 2016, 52, 7020. [9] INVITED AND PROMOTED LECTURES Sauvée, C., M. Rosay, G. Casano, et al., Angew. Chemie Int. Ed. 2013, 52, 10858. [10] Geiger, M. A., A. P. Jagtap, M. Kaushik, et al. , Chem Eur J, 2018, 24, 13485. [11] POSTERS Stevanato, G., G. Casano, D. J. Kubicki, et al. , J. Am. Chem. Soc. 2020 , 142, 16587. ADVERTISERS Acknowledgements: The National High Magnetic Field laboratory (NHMFL) is funded by the National Science Foundation Division of Materials Research AUTHOR INDEX (DMR-1644779) and the State of Florida. A portion of this work was supported by the NIH P41 GM122698. PT021 PARALLEL SESSIONS / PROMOTED TALK / HYPERPOLARIZATION 76 GEORGES MENZILDJIAN1, BDPA-NITROXIDE BIRADICALS FOR EFFICIENT DNP ALICIA LUND1, GABRIELE ENHANCED SOLID-STATE NMR AT HIGH FIELDS AND ELEVATED STEVANATO2, MAXIM YULIKOV3, GILLES TEMPERATURE CASANO4,GUNNAR JESCHKE3, DAVID GAJAN1, LYNDON Over the last decade, dynamic nuclear polarization (DNP) has be- In this context, we will first present a new family of BDPA-nitroxide EMSLEY2, MORENO LELLI5, come a cornerstone technique to circumvent the inherent sensitiv- radicals based on a 5-membered ring nitroxide. The performances OLIVIER OUARI4, ity limitations of solid-state magic angle spinning (MAS) NMR. At of the best radical of the series will be presented at 18.8 and 21.1 T ANNE LESAGE1 very high magnetic fields (typically 18.8 T and above) however, the and compared to the HyTEK series. While the DNP efficiency of the 1 gold standard binitroxide polarizing agents developed at 9.4 T and Hy-5 radicals do not exceed those reported for HyTEK polarizing High-Field NMR Center, University of Lyon, Lyon, France yielding 250-fold signal enhancements fail to produce such signif- agents, this study provides new clues to fine-tune and modulate the 2Institut des Sciences et Ingénierie Chimiques, icant gains. structure of hybrid radicals. EPFL, Lausanne, Switzerland 3Department of Chemistry and Applied A new class of hybrid polarizing agents based on BDPA-nitroxide We will then discuss the performance of BDPA-nitroxide radicals Biosciences, ETH Zürich, Zürich, Switzerland 4 biradicals, HyTEKs, was introduced in 2018 by Wisser and co-work- at elevated temperatures. One of the main limitation of DNP MAS Institut de Chimie Radicalaire Aix Marseille Univ, Marseille, France ers [1] and shown to successfully circumvent the limitations of bini- NMR is its limited applicability to cryogenic temperatures. By in- 5Center of Magnetic Resonance (CERM), troxides at high fields, yielding enhancements as high as 180-fold at corporating BDPA or TEKPol in ortho-tertphenyl (OTP), a rigid ma- University of Florence, Florence, Italy 18.8 T spinning at 40 kHz MAS in a 1.3 mm probe, or more recently trix with a glass transition temperature (T ) about 243 K, Lelli and g 200-fold at 21.1 T spinning at 65 kHz in a 0.7 mm probe [2]. These co-workers were able to improve the performance of these radicals hybrid radicals take advantage of the narrow (and therefore easy to at elevated temperatures [3]. In this work, we will show that by for- INTRODUCTION saturate) isotropic BDPA EPR line, strong electron-electron magnet- mulating HyTEK2 in OTP enhancements as high as 65 at 230 K at SPONSORS ic interactions and long electronic relaxation times thanks to the 18.8 T were maintained. The DNP performance of this system as a PROGRAM functionnalization of the nitroxide moiety. function of temperature will be presented and discussed in the light PROGRAM — BY DAY of variable temperature EPR relaxation measurements and spin dy- TOPICS namics simulations using the SPINACH software [4]. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] D. Wisser et al., J. Am. Chem. Soc. 2018, 140, 13340-13349. [2] P. Berruyer et al., J. Phys. Chem. Lett. 2020, 11, 8386-8391. [3] M. Lelli et al., J. Am. Chem. Soc. 2015, 137, 14558-14561. AUTHOR INDEX [4] H. J. Hogben et al., J. Magn. Reson. 2011, 208, 179-194. PT022 PARALLEL SESSIONS / PROMOTED TALK / HYPERPOLARIZATION 77 VENKATA SUBBARAO TPPM DNP REDROUTHU, XIAOXUN CHEN, In pulsed DNP, microwave irradiation is applied to an NMR sample In Off-resonance NOVEL, a low electron nutation frequency is com-SANJAY VINOD KUMAR, in the form of a pulse sequence with the aim to enhance the bulk pensated by irradiating off-resonance. In TOP DNP, which consists GUINEVERE MATHIES nuclear polarization, and thereby the sensitivity of the NMR exper- of a train of pulses and delays ( ), a com- iment. This relatively new way of doing DNP may have several ad- bination of the modulation frequency ( ) and the Department of Chemistry, University of Konstanz, Konstanz, Germany vantages over existing, classical forms of DNP, which all rely on con- effective frequency of the electron spins under the influence of the tinous microwave irradiation. These include an effective Hamilto- pulse train ( ) must equal the nuclear nian that does not depend on the static magnetic field, thus holding Larmor frequency, which further increases the possibilities when the promise of a field-independent DNP efficiency, and the ability the electron nutation frequency is low. to generate DNP at a low duty cycle, which reduces sample heating, Here we introduce two new DNP pulse sequences: TPPM DNP (Two particularly at the highest magnetic fields. Pulse Phase Modulation, ) The first DNP pulse sequence, NOVEL (Nuclear spin Orientation Via and XiX DNP (X-inverse-X, ), Electron spin Locking),[1] dates back to the 1980s. NOVEL works named after the analogous, well-known heteronuclear decoupling well in chemical systems compatible with DNP/MAS NMR of bio-sequences. For both sequences, the matching condition is the same molecular systems, e.g. trityl in DNP juice, but the sequence is tech- as for TOP DNP, but because the trajectories of the electron spins dif- nically challenging to implement at high magnetic fields, because fer, will be different, as well as the scaling factors. during the contact period the electron nutation frequency ( ) In experiments at Q band (51 MHz/1.2 T/34 GHz), using a nutation must match the 1H Larmor frequency ( ). Fortunately, in recent frequency of 18 MHz, we have found that XiX DNP is superior to TOP INTRODUCTION years two new DNP sequences have been introduced, for which the DNP. Numerical simulations of polarization transfer in an ensemble SPONSORS requirement on the electron nutation frequency is considerably of dipolar coupled e--1H spin systems were set up with Spinach[4] PROGRAM relaxed. These are: Off-resonance NOVEL,[2] with the matching and were extremely helpful for screening the XiX and TPPM match- PROGRAM — BY DAY condition ing conditions. TOPICS PRIZE LECTURES PLENARY LECTURES and TOP (Time-Optimized Pulsed) DNP,[3] with the matching TUTORIAL LECTURE condition INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] A. Henstra, P. Dirksen, J. Schmidt, W. Th. Wenckebach, J. Magn. Reson. 1988, 77, 389-393. [2] S. K. Jain,* G. Mathies,* R. G. Griffin, J. Chem. Phys. 2017, 147, 164201. [3] K. O. Tan, AUTHOR INDEX C. Yang, R. T. Weber, G. Mathies, R. G. Griffin, Sci. Adv. 2019, 5, eaav6909. [4] H. Hogben, M. Krzystyniak, G. T. P. Charnock, P. J. Hore, I. Kuprov, J. Magn. Reson. 2013, 208, 179–194. PT023 PARALLEL SESSIONS / PROMOTED TALK / HYPERPOLARIZATION 78 AARON HIMMLER1, IN-SITU EPR AND DNP-ENHANCED NMR OF SILICON MOHAMMED M. NANO PARTICLES ALBANNAY1,2, GEVIN VON WITTE1,2, Nuclear magnetic resonance (NMR) and electron paramagnetic resonance (EPR) are two established methods for MATTHIAS ERNST1 studying the structure of engineered materials such as silicon-based micro- and nano- particles [1]. Direct 29Si NMR 1ETH Zurich, Laboratory of detection is challenging due to its low thermal polarization; however low-temperature (<4 K) dynamic nuclear po-Physical Chemistry, Zurich, larization (DNP) can slowly enhance sensitivity. Albeit, the endogenous free-radicals (lattice defects) driving the Switzerland 2University and ETH Zurich, DNP process are in low concentrations [2]. Consequently, EPR sensitivity is low thus requiring intense microwave Institute for Biomedical irradiation and frequency modulation to sufficiently saturate the target electron transition. But unfortunately, sol-Engineering, Zurich, Switzerland id-state microwave sources are inefficient at high frequencies. We present a technical solution for NMR and EPR detection to study the DNP efficiency of crystalline silicon particles in our 7 T DNP polarizer [3]. The over-sized steel WR28 waveguide coupling microwaves from the source (197 GHz Tx, Virginia Diodes) is electroplated with a layer of gold, palladium and finally silver to reduce transmission losses at 197 GHz. The method involves reducing metallic cations on the waveguide walls using a specially constructed plating anode that holds the desired electrolyte. Unlike traditional electroplating methods, the electrolyte solution doesn’t require heating nor intermittent pH and surface layer treatment, allowing for use on a regular lab bench. To facilitate EPR detection, a 750-turn solenoid coil fabricated from 0.1 mm diameter copper wire is fitted onto the DNP. The 29Si DNP spectra and longitudinal-EPR for the crystalline silicon particles (P28A002, Alfa Aesar), are illustrated in Fig 1B. Transmission loss was characterized on the bench using a diode detector (WR5.1ZBD-F, Vir- INTRODUCTION ginia Diodes). Plating the 655 mm long waveguide resulted in a decrease of the loss by 3.1 dB. Testing eight months SPONSORS later yielded similar results suggesting good plate adhesion during electroplating and low corrosion. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] E. Tasciotti, et al. , Nat. Nanotechnol. 2008, 3, 151–157. [2] Kwiatkowski G, et al. J. Phys. Chem. C, 2018. [3] Jähnig F et al. , Phys. Chem. Chem. Phys. AUTHOR INDEX 2017. IN019 PARALLEL SESSIONS / INVITED SPEAKER / METHODS DEVELOPMENT 79 AMRIT VENKATESH1,2, INDIRECT DETECTION METHODS FOR THE RAPID MEASUREMENT BENJAMIN A. ATTERBERRY1,2, OF ISOTROPIC AND ANISOTROPIC 195PT CHEMICAL SHIFTS IN SCOTT L. CARNAHAN1,2, LUKAS ROCHLITZ3, HETEROGENEOUS CATALYSTS PENGXIN LIU3, ZHUORAN WANG4, Platinum is commonly found in heterogeneous catalysts in the form pulses can also be implemented into 1H{195Pt} symmetry-based ro- GEORGES MENZILDJIAN4, of dispersed atoms/ions, immobilized organometallic compounds tational echo double resonance (S-REDOR) experiments to increase ANNE LESAGE4, and metal nanoparticles. 195Pt is a 33% naturally abundant spin-1/2 dipolar dephasing and facilitate more accurate measurement of CHRISTOPHE COPÉRET3, nucleus with a gyromagnetic ratio close to that of 13C, making 195Pt 1H-195Pt dipolar coupling constants and internuclear distances,[4] AARON J. ROSSINI1,2 solid-state NMR is an appealing technique to study the molecular including in model catalysts.[3] We then present a novel fast MAS, 1 structure of platinum catalysts. However, 195Pt solid-state NMR ex- indirect detection approach to rapidly measure 195Pt CSA. The high Iowa State University, Department of Chemistry, Ames, IA, USA periments are often challenging, if not impossible, because 195Pt power 195Pt pulses used in D-HMQC/S-REDOR are replaced with 2US DOE Ames Laboratory, Ames, IA, USA chemical shift anisotropy (CSA) can easily exceed 8000 ppm. Fur- long-duration, low-power, sideband-selective excitation/saturation 3Department of Chemistry and Applied thermore, many heterogeneous catalysts feature low metal load- pulses. The 195Pt spinning sidebands can then be mapped out by Biosciences, ETH Zürich, Zürich, Switzerland 4 High-Field NMR Center of Lyon, CNRS, ENS ings on the order of 0.5-4 wt.%. We have previously shown that fast varying the offset of the 195Pt sideband selective pulses, enabling the Lyon, UCB Lyon, Villeurbanne, France MAS, 1H{195Pt} dipolar heteronuclear multiple quantum correlation rapid measurement of the chemical shift tensor parameters with (D-HMQC) experiments can be used for the sensitive and high-res-high sensitivity. Using these approaches, we were able to measure olution detection of isotropic and anisotropic 195Pt chemical shifts. 195Pt CSA exceeding 8000 ppm in only a few minutes for pure com- [1-3] In this presentation we describe new solid-state NMR methods pounds and a few hours for a Pt compound grafted on silica with that enhance sensitivity and facilitate high-resolution detection of ca. 4 wt.% Pt loading. In comparison, DNP-enhanced wideline 195Pt INTRODUCTION wideline 195Pt solid-state NMR spectra. CPMG experiments required many days of acquisition to measure SPONSORS the CSA. Therefore, these indirect detection methods pave the way PROGRAM First, we will show new t -noise eliminated (TONE) D-HMQC that 1 for the analysis of complex Pt-based heterogeneous catalysts with PROGRAM — BY DAY provide order of magnitude gains in sensitivity by suppressing t -1 low loadings. TOPICS noise.[4] The TONE D-HMQC experiments exploit short high-pow- PRIZE LECTURES er adiabatic pulses (SHAP) for broadband inversion of 195Pt. SHAP PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS [1] Rossini A.J., Hanrahan M.P., Thuo M., Physical Chemistry Chemical Physics, 2016, 18, 25284-25295. [2] Perras F.A., Venkatesh A., Hanrahan M.P., Goh T.W., Huang W., Rossini A.J., Pruski M. J. ADVERTISERS Magn. Reson. , 2017, 276, 95-102. [3] Venkatesh, A., Lund, A., Rochlitz, L., Jabbour, R., Gordon, C. P., Menzildjian, G., Viger-Gravel, J., Berruyer, P., Gajan, D., Copéret, C., Lesage, A., Rossini, A. J., AUTHOR INDEX J. Am. Chem. Soc. 2020, 142, 18936-18945. [4] Venkatesh A., Perras F.A., Rossini A.J., J. Magn. Reson. 2021, 106983. IN020 PARALLEL SESSIONS / INVITED SPEAKER / METHODS DEVELOPMENT 80 NINA ELENI CHRISTOU, LIGHT NMR: A HIGH-RESOLUTION TOOL FOR THE MECHANISTIC DOMINIQUE BOURGEOIS, UNDERSTANDING OF PHOTOPHYSICAL PROPERTIES OF BERNHARD BRUTSCHER Institut de Biologie Structurale, FLUORESCENT PROTEINS University Grenoble Alpes, CEA, CNRS, Grenoble, France Fluorescent proteins with distinct photo-transformation properties sample illumination at 3 different wavelengths (405, 488, and 561 are crucial for a wide range of applications in advanced fluores- nm) that can be synchronized with NMR pulse sequences. A set of cence microscopy and biotechnology. However, using photo-trans- multidimensional and multinuclear (1H, 13C, 15N) NMR experiments, formable fluorescent proteins (PTFPs) remains highly challenging including 15N relaxation, H/D exchange, pH titration measure-because environmental conditions such as pH, temperature, salt, ments, light-driven EXSY, and specific chromophore experiments, oxygen level, reducing agents, and viscosity affect their brightness, were setup to build an “NMR investigation toolbox” for fluorescent photostability and photo-transformation landscape. Rational fluo-proteins [1]. Studying rsFolder, a reversibly switchable green fluo- rescent protein design currently exploits the mechanistic informa- rescent protein, we have identified four distinct configurations of tion available from structural studies, mainly X-ray crystallography, its p-HBI chromophore, corresponding to the cis and trans isomers, in order to design new PTFP variants with improved properties for with each one either protonated (neutral) or deprotonated (anionic) particular applications. Yet, protein crystals only grow under en-at the benzylidene ring. The relative populations and interconver- vironmentally-restrictive crystallization conditions and crystallo- sion kinetics of these chromophore species depend on sample pH graphic data, often recorded at cryogenic temperatures, lack im- and buffer composition that alter in a complex way the strength of portant information on structural heterogeneity and conformation- H-bonds that contribute in stabilizing the chromophore within the al dynamics. Thus crystallographic structures may poorly account protein scaffold [2], and determine its fluorescence and switching INTRODUCTION for photophysical properties observed in biological samples at quantum yields. SPONSORS physiological temperature. Multidimensional solution NMR spec- PROGRAM Light-NMR spectroscopy provides a powerful tool to investigate the troscopy provides an unique tool to investigate at atomic resolution PROGRAM — BY DAY effects of environmental conditions or protein mutations on chro- the conformational dynamics of PTFPs in different photo-stationary TOPICS mophore state populations and dynamics, and to correlate them states, under a variety of physicochemical conditions, and over a PRIZE LECTURES with altered photophysical properties as a prerequisite for rational wide range of time scales. PLENARY LECTURES design of phototransformable fluorescent protein variants with im- TUTORIAL LECTURE We have set up a portable NMR in-situ illumination device that is proved properties. INVITED AND PROMOTED LECTURES compatible with high-field NMR spectrometers, allowing NMR POSTERS ADVERTISERS AUTHOR INDEX References: [1] Christou et al., Biophys. J. 2019, 117, 2087-2100. [2] Christou et al., J. Am. Chem. Soc. 2021, https://doi.org/10.1021/jacs.1c02442. IN021 PARALLEL SESSIONS / INVITED SPEAKER / METHODS DEVELOPMENT 81 OTTO MANKINEN1, ULTRAFAST DIFFUSION EXCHANGE SPECTROSCOPY VLADIMIR V. ZHIVONITKO1, ANNE SELENT1, Relaxation and diffusion NMR (called Laplace NMR, LNMR) experiments offer insights into molecular dynamics. SARAH MAILHIOT1, Multidimensional approach improves the resolution of LNMR. Furthermore, it enables one to correlate diffusion SANNA KOMULAINEN1, and relaxation parameters and study exchange phenomena through the relaxation or diffusion contrast. Multidi- NØNNE L. PRISLE2, mensional LNMR experiments are, however, very time consuming, as the experiment must be repeated many times SUSANNA AHOLA1, Y with an incremented evolution time delay or gradient strength. ASHU KHARBANDA1, Recently, we have demonstrated that multidimensional LNMR can be accelerated by one to three orders of mag- MATEUSZ URBAŃCZYK1,3, nitude by spatial encoding [1,2], which was originally exploited in ultrafast (UF) NMR spectroscopy [3]. We call the MIKKO KETTUNEN4, method UF LNMR. We have exploited UF LNMR in the investigation of porous media, chemical analysis, identifica- VILLE-VEIKKO TELKKI1 tion of intra- and extracellular metabolites as well as mobile NMR [1,2,4-7]. The single-scan approach also signifi-1NMR Research Unit, University of Oulu, Oulu, cantly facilitates the use of hyperpolarization to boost sensitivity by many orders of magnitude [2,4,7]. Finland 2Center for Atmospheric Research, University This talk concentrates on the description of a novel ultrafast diffusion exchange spectroscopy (UF DEXSY) method of Oulu, Oulu, Finland 3 [8]. Conventional DEXSY is very time consuming, as both the indirect and direct dimensions are collected point by Centre of New Technologies, University of Warsaw, Warsaw, Poland point. In the UF DEXSY, complete data is measured in a single scan by exploiting spatial encoding in the indirect 4A.I. Virtanen Institute for Molecular Sciences, dimension and CMPG based detection accompanied by gradients, which serve both as read and diffusion gradients. University of Eastern Finland, Kuopio, Finland We have exploited the method in studying exchange phenomena in surfactant solutions relevant to aerosol re- search. Furthermore, we have preliminary data showing that the experiments are feasible also with a low-field single-sided NMR spectrometer (NMR-MOUSE), and the significant sensitivity boost provided by dissolution dynamic INTRODUCTION nuclear polarization (dDNP) method allows single scan analysis of molecular exchange in a yeast cell solution. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] S. Ahola, V.-V. Telkki, ChemPhysChem, 2014, 15, 1687. [2] Ahola et al., Nat. Commun., 2015, 6, 8363. [3] L. Frydman et al., Proc. Natl. Acad. Sci. INVITED AND PROMOTED LECTURES USA, 2002, 99, 15858. [4] O. Mankinen et al., Micropor. Mesopor. Mat., 2018, 269, 75. [5] G. Zhang et al., Anal. Chem., 2018, 90, 11131. [6] J.N. King et al., Angew. POSTERS Chem. Int. Ed., 2016, 55, 5040. [7] J.N. King et al., Chem. Sci., 2018, 9, 6143. [8] O. Mankinen et al., Nat. Commun., 2020, 11, 3251. ADVERTISERS Acknowledgements: The authors gratefully acknowledge funding from the European Research Council under Horizon 2020, projects Ultrafast Laplace NMR AUTHOR INDEX (grant agreement no. 772110). PT024 PARALLEL SESSIONS / PROMOTED TALK / METHODS DEVELOPMENT 82 JAVIER A. ROMERO1, ACCELERATED ACQUISITION OF 2D TITRATION DATA WITH NON- EWA K. NAWROCKA1,2, STATIONARY COMPLEMENTARY NON-UNIFORM SAMPLING ALEXANDRA SHCHUKINA3, FRANCISCO J. BLANCO4, (NOSCO-NUS) TAMMO DIERCKS5, KRZYSZTOF KAZIMIERCZUK1 The versatility of NMR and its broad applicability to several stages ligand-to-protein ratio is sampled for a unique set of t times. A sep-1 1 in the drug discovery process is well known and generally consid- arate reference spectrum of the protein before adding ligand is also Center of New Technologies, University of Warsaw, Warsaw, Poland, ered one of the major strengths of NMR. Indeed, NMR is the only recorded. The various NUS 2D time-domain data of a titration series 2Faculty of Chemistry, University of Warsaw, biophysical technique which can detect and quantify molecular can be combined and compressed to yield a single 2D correlation Warsaw, Poland, 3 interactions, and at the same time provide detailed structural in- spectrum. Signals that shift strongly (i.e. show large CSP) appear Faculty of Chemistry, Biological and Chemical Research Centre, UW, Warsaw, formation with atomic-level resolution [1]. This powerful NMR ap- smeared out and broadened due to superposition of their variable Poland, plication requires long measurement times to record well-resolved positions in the different underlying 2D spectra of the titration se-4CIB-CSIC, Structural and Chemical Biology Department, Madrid, Spain, heteronuclear 2D correlation spectra of the protein, where the sig- ries. Such NMR signals can be modeled as non-stationary signals [3]. 5CIC bioGUNE, Parque Tecnológico Bizkaia, nals of amino acids affected by ligand binding gradually shift or dis- The idea powering NOSCO-NUS is to use the traceable dependence Derio, Spain appear with increasing ligand concentration. The time demand is of t times with each titration point in the series to treat the smeared 1 heightened by the need to record full titration series to determine signals, refocusing them into the sharp reference signal with a con-the affinity constant and quantify the chemical shift perturbation comitant increase in intensity. This is achieved by multiplying each (CSP) amplitude. selected NMR signal by a synthetic signal which obeys the same single-site binding model followed by the shifting peaks. Accurate We propose a method for complementary non-uniform sampling binding parameters (affinity constant and CSP in both dimensions) INTRODUCTION (NUS) of entire titration series of heteronuclear 2D correlation are found when peak height difference between reference and cor- SPONSORS spectra within less time than even required for a single conven- rected signals is minimal. The precision of NOSCO-NUS is shown to PROGRAM tional spectrum [2]. For this we sample the indirect t1 dimension be comparable to conventional methods. PROGRAM — BY DAY as in a standard NUS experiment, yet with complementary sam- TOPICS pling schedules for the different titration points such that each PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] M. P. Williamson, Prog. Nucl. Magn. Reson. Spectrosc. 2013, 73, 1-16 [2] J. A. Romero, E. K. Nawrocka, A. Shchukina, F. J. Blanco, T. Diercks, K. Kazimierczuk, Angew. Chem. Int. Ed. POSTERS 2020, 132, 23702-23705 [3] D. Gołowicz, P. Kasprzak, V. Orekhov, K. Kazimierczuk, Prog. Nucl. Magn. Reson. Spectrosc. 2020, 116, 40-45 ADVERTISERS Acknowledgements: The authors would like to thank the National Science Centre of Poland for its support in the form of a HARMONIA grant (2017/26/M/ST4/01053) and the Foundation for AUTHOR INDEX Polish Science for its support under the FIRST TEAM programme, co-financed by the European Union under the European Regional Development Fund no. POIR.04.04.00-00-4343/17-00. PT025 PARALLEL SESSIONS / PROMOTED TALK / METHODS DEVELOPMENT 83 SEBASTIAN TASSOTI1, MONITORING FAST CHEMICAL PROCESSES SIMON GLANZER1, BY REACTION-INTERRUPTED NMR SPECTROSCOPY EDUARD STADLER2, GABRIEL E. WAGNER1, NMR spectroscopy is a powerful, widely used tool for the investi- of a selected atom to follow its conversion in the course of a fast re- GEORG GESCHEIDT2 AND gation of chemical reactions. It has been used to study kinetics of action. The speed of recording irreversible reactions by in situ NMR KLAUS ZANGGER1 unstable reaction intermediates, mechanisms in organic chemistry can be significantly enhanced by combining rapid injection devices 1University of Graz, Institute of and for process control in so-called inline/online NMR [1]. A further with very fast data acquisition schemes, which rely on relaxation Chemistry, Graz, Austria 2 advantage compared to other methods is that there is no necessity independent observation of individual NMR tube slices by spatially Graz University of Technology, Institute of Physical and Theoretical Chemistry, for a prior calibration, as the integral of a signal is directly propor- selective excitation [3]. Graz, Austria tional to the quantity of the material present. NMR Spectroscopy is In contrast to ExTra NMR, reversible photoinduced reactions can generally used to study stable molecules under equilibrium condi-be investigated by an light pulse interrupted NOESY/EXSY exper- tions. Here we present a suite of experiments, that we developed in iment. Irradiation during the mixing period of a modified EXSY order to study fast reversible and irreversible reactions, which are experiment leads to additional peaks in the resulting two-dimen-ongoing during the pulse-sequence. sional spectra [4]. The photo-induced exchange of magnetization Despite recent technological and methodogical developments occurring in photo-switchable (Z) and (E) forms of azo compounds to study on-going reactions, tracing the fate of individual atoms provides information on the corresponding dynamic equilibria. We during an irreversible chemical reaction is still a challenging and report on the dependence of the diagonal-to-cross-peak ratio on elaborate task. Reaction-interrupted excitation transfer (ExTra) concentration, light intensity and mixing time. Reversible chem-NMR provides a selective tracking of resonances from atoms, which ical processes can also be investigated by a 2D experiment, which INTRODUCTION undergo chemical conversion [2]. We show that reactions, triggered allows the monitoring of CEST between any two signals at once. SPONSORS either by rapid mixing or by photo-excitation can be conveniently This is achieved by a combination of slice-selective excitation with PROGRAM followed at a sub-second time scale using standard NMR equip- obersavation during a weak pulsed field gradient. PROGRAM — BY DAY ment. In ExTra NMR we use the selectively inverted magnetization TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] D. A. Foley, J. Wang, B. Maranzano, M. T. Zell, B. L. Marquez, Y. Xiang and G. L. Reid, Anal. Chem., 2013, 85, 8928–8932. [2] G.E. Wagner, S. Tassoti, S. Glanzer, E. Stadler, R. INVITED AND PROMOTED LECTURES Herges, G. Gescheidtc and K. Zangger, Chem. Commun. 2019, 55, 12575-12578. [3] G. E. Wagner, P. Sakhaii, W. Bermel and K. Zangger, Chem. Commun., 2013, 49, 3155-3157. [4] E. Stadler, S. POSTERS Tassoti, P. Lentes, R. Herges, T. Glasnov, K. Zangger and G. Gescheidt, Anal. Chem., 2019, 91, 11367–11373. ADVERTISERS Acknowledgements: Financial support by the Austrian Science Foundation (FWF) (Project Number P-30230) and through the Doktoratskolleg Molecular Enzymology (W901) is gratefully AUTHOR INDEX acknowledged. We also thank the interuniversity program BioTechMed and the research focus BioHealth, for financial contributions. PT026 PARALLEL SESSIONS / PROMOTED TALK / METHODS DEVELOPMENT 84 IVAN ZHUKOV1, TOTAL CHEMICAL SHIFT CORRELATION AMONG ALEXEY KIRYUTIN1, ALL MAGNETIC NUCLEI BY ISOTROPIC MIXING IN FABIEN FERRAGE2, GERD BUNTKOWSKY3, ZERO- TO ULTRA-LOW FIELD GEOFFREY BODENHAUSEN2, ALEXANDRA YURKOVSKAYA1, Any two-dimensional NMR pulse sequence contains the same coil system located in a µ-metal magnetic shield. Armed with such AND KONSTANTIN IVANOV1,† functional intervals, or principal building blocks: (i) preparation; unique equipment, we realized an attractive idea to fulfil the condi-1 (ii) evolution; (iii) mixing; (iv) detection [1]. In a well-known TOCSY tion of strong coupling for any heteronuclei by utilizing zero- to ul-International Tomography Center and Novosibirsk State University, Novosibirsk, experiment [2], which highlights the network of coupled nuclei, tra-low field while keeping high-resolution NMR detection at high Russia the mixing block is given by a sufficiently strong spin-lock pulse. field. Here we present this novel approach to obtain a 2D chemical 2École Normale Supérieure, Département In this experiment, an isotropic mixing of polarization due to scalar shift correlation spectrum with polarization transfer between het- de Chimie, and CNRS, Paris, France 3Technical University of Darmstadt, I spin-spin coupling between locked nuclear spins occurs during the eronuclei named ZULF-TOCSY [5]. The mixing block of ZULF-TOCSY nstitute of Inorganic and Physical spin-locking pulse application. The heteronuclear version of TOCSY contains an interval of isotropic mixing at zero- to ultra-low field. Chemistry, Darmstadt, Germany known as HEHAHA (Heteronuclear Hartmann-Hahn) [3] relies on This experiment in principle allows observing all possible correla-polarization transfer between heteronuclei once Hartmann-Hahn tions between signals of all magnetic nuclei with different gyro-conditions are met. However, in practice, the application of the HE- magnetic ratios. This unique feature assigns the application of the HAHA pulse sequence is limited by technical problems since strong ZULF-TOCSY experiment for the straightforward determination of radiofrequency irradiation is necessary to cover the entire distribu- mixture components. In addition, ZULF-TOCSY allows rendering tion of heteronuclei chemical shifts. sequential assignments in peptides at natural abundance then no amide protons are present using a single 2D spectrum [6]. The iso- INTRODUCTION A few years ago, a cost-efficient shuttling add-on compatible with tropic mixing at zero- to ultra-low field can be used to construct SPONSORS commercial NMR spectrometer was built in our lab [4]. Ultra-wide more sophisticated NMR pulse sequences in the future. PROGRAM field variation from 5 nT to 9.4 T was made possible by sample posi- PROGRAM — BY DAY tioning along the bore of NMR magnet compiled with electromagnet TOPICS PRIZE LECTURES PLENARY LECTURES † Konstantin Ivanov passed away on March 5th, 2021 at the age of 44 years as a consequence of Covid-19. TUTORIAL LECTURE References: [1] R.R. Ernst, G. Bodenhausen, and A. Vokaun Principles of Nuclear Magnetic Resonance in One and Two Dimensions, Clarendon Press, Oxford, 1987. [2] L. Braunschweiler, R. R. INVITED AND PROMOTED LECTURES Ernst, J. Magn. Reson. 1983, 53, 521-528. [3] G. A. Morris, A. Gibbs, Magn. Reson. Chem. 1991, 29, 83-87. [4] I.V. Zhukov, A.S. Kiryutin, A.V. Yurkovskaya, Y. A. Grishin, H.-M. Vieth, and K.L. POSTERS Ivanov, Phys. Chem. Chem. Phys. 2018, 20, 12396-12405. [5] I.V. Zhukov, A.S. Kiryutin, F. Ferrage, G. Buntkowsky, A.V. Yurkovskaya, and K.L. Ivanov, J. Phys. Chem. Lett. 2020, 11, 7291-7296. [6] ADVERTISERS A.S. Kiryutin, I.V. Zhukov, F. Ferrage, G. Bodenhausen, A.V. Yurkovskaya, K.L. Ivanov, Phys. Chem. Chem. Phys. 2021, 23, 9715-9720. AUTHOR INDEX Acknowledgements: This work was supported by Russian Foundation for Basic Research (#19-29-10028). PT027 PARALLEL SESSIONS / PROMOTED TALK / METHODS DEVELOPMENT 85 PAWEŁ KASPRZAK1, POISON-GAP SAMPLING EXPLAINED MATEUSZ URBAŃCZYK2, KRZYSZTOF KAZIMIERCZUK2 Non-uniform sampling (NUS) became a popular way of accelerating time-consuming multidimensional NMR 1 experiments. Among various non-conventional sampling schemes, the Poisson-gap (PG) schedules [1,2] became Faculty of Physics, University of Warsaw, Warsaw, Poland; particularly popular, especially when combined with compressed sensing (CS) reconstruction of the missing data 2Centre of New Technologies, University of points [3]. However, the effectiveness of PG is based mainly on practical experience and has not been so far ex-Warsaw, Warsaw, Poland plained based on the CS theory. Moreover, there is an apparent contradiction between the observed potential of PG and CS theory which states, that a “flat” pseudo-random generator is the most optimal way of generating sampling schedules. In this presentation, we explain why and when PG reveals its superior features in NMR spectroscopy. We support our theoretical considerations with simulations and analysis of experimental data from BMRB. The latter reveals (so far unnoticed) feature of NMR spectra explaining the success of PG and related schedules. We name this feature the “clustered” sparsity. In other words, the NMR spectra are not only sparse, but the peaks often form groups in the indirect dimension. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Kazimierczuk, K., Zawadzka, A., Koźmiński, W., J. Magn. Reson. 2008, 192(1), 123–130. [2] Hyberts, S. G., Takeuchi, K., & Wagner, G., J. Am. Chem. INVITED AND PROMOTED LECTURES Soc., 2010, 132(7), 2145–2147. [3] Hyberts, S. G., Milbradt, A. G., Wagner, A. B., Arthanari, H., Wagner, G., J. Biomol. NMR 2012, 52(4), 315–327. POSTERS Acknowledgements: The authors would like to thank the National Science Centre of Poland for its support in the form of a HARMONIA grant (2017/26/M/ ADVERTISERS ST4/01053) and the Foundation for Polish Science for its support under the FIRST TEAM programme, co-financed by the European Union under the European AUTHOR INDEX Regional Development Fund no. POIR.04.04.00-00-4343/17-00. PT028 PARALLEL SESSIONS / PROMOTED TALK / METHODS DEVELOPMENT 86 JIAFEI MAO1, ROOM-TEMPERATURE DNP NMR SPECTROSCOPY OF DANHUA DAI2, SMALL BIOLOGICAL MOLECULES IN WATER VASYL DENYSENKOV2, XIANWEI WANG3,4, Nuclear magnetic resonance (NMR) spectroscopy is a powerful and popular technique for probing molecular struc- YIWEI LIU3, XIAO HE3,5, tures, dynamics and chemical properties. However the conventional NMR spectroscopy is bottlenecked by its low THOMAS PRISNER2 sensitivity. Dynamic nuclear polarization (DNP) boosts NMR sensitivity by orders of magnitude and resolves this 1Goethe University Frankfurt, Institute limitation. In liquid-state this revolutionizing technique is still restricted to a few specific model molecules in or-of Biophysical Chemistry and Center ganic solvents. Here we present that, for the first time, a full scheme of small biological molecules, ranging from car-for Biomolecular Magnetic Resonance, Frankfurt am Main, Germany bohydrates to amino acids, have been hyperpolarized efficiently in water directly at room temperature and at high-2Goethe University Frankfurt, Institute field. A trend between observed ODNP enhancement factor and paramagnetic shifts have been revealed, which of Physical and Theoretical Chemistry instructs us to revisit paramagnetic NMR literatures and to discover new class of molecules (heterocyclics) that and Center for Biomolecular Magnetic Resonance, Frankfurt am Main, Germany can be hyperpolarized by Overhauser DNP. The QM/MM MD simulation underscores the dynamic intermolecular 3East China Normal University, School of hydrogen bonds as the driving force for Overhauser DNP. Our work reconciles DNP, paramagnetic NMR and com- Chemistry and Molecular Engineering, Shanghai, China putational chemistry, illuminates unexplored molecular space for modern liquid-state DNP NMR spectroscopy, and 4Zhejiang University of Technology, College reveals new molecular and chemical mechanism of Overhauser DNP in liquids. of Science, Hangzhou, Zhejiang, China 5NYU-ECNU Center for Computational Chemistry at NYU Shanghai, Shanghai, China INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PT029 PARALLEL SESSIONS / PROMOTED TALK / METHODS DEVELOPMENT 87 KATHRIN AEBISCHER1, RADIAL RADIO-FREQUENCY FIELD INHOMOGENEITY IN ZDENĚK TOŠNER2, MAS SOLID-STATE NMR EXPERIMENTS MATTHIAS ERNST 1Physical Chemistry, ETH Zürich, Radiofrequency-field (rf) inhomogeneity is a prevalent problem in weaker sidebands due to amplitude modulations are observed Zürich, Switzerland 2 NMR experiments that often leads to the deterioration of the per- around the nominal rf amplitude. The intensity of such sidebands Department of Chemistry, Charles University, Prague, Czech Republic formance of pulse sequences. The extent of the inhomogeneity is can help to characterize the rf-field modulations and thus give in-largely determined by the coil geometry and different coil designs sight into the radial contribution to the rf-field inhomogeneity for have been proposed to improve rf homogeneity. In solid-state NMR the MAS probe used. For polarization transfer sequences such as probes, solenoid coils wound around the rotor inside the MAS sta-Hartmann-Hahn cross polarization, REDOR and symmetry-based C tor are still most commonly used, even though they suffer from sequences only minor effects of the rf amplitude and phase modu-comparatively large rf inhomogeneity. The spatial rf-field distribu- lations were observed that should be of no consequence for experi- tion over the sample volume in such MAS probes is mainly charac- mental implementations. In simulated homonuclear spectra under terized by the distribution along the rotor axis. However, significant FSLG decoupling, significant line broadening occurred when MAS radial contributions are also present [2]. Under MAS, these radial modulation of the rf amplitude was taken into account. Floquet contributions lead to a time-dependent modulation of the rf am-analysis of the effective Hamiltonian up to second order revealed plitude and phase experienced by a spin packet [3]. In contrast to that this broadening is most likely due to the re-introduction of the static part of the rf inhomogeneity, the effects of these time-de-homonuclear dipolar coupling terms in the first-order effective pendent modulations are not well-studied. Here we present an in- Hamiltonian. However, no experimental characterization of this vestigation of the influence of such MAS-induced rf amplitude and additional line broadening was possible, as experimentally ob- INTRODUCTION phase modulations on building blocks commonly encountered in tained line widths were not decoupling limited. SPONSORS solid-state NMR pulse schemes. Potential effects were studied using Overall, our findings suggest that the effects of the radial contri- PROGRAM numerical simulations and analytical approaches based on Floquet bution to the rf-field inhomogeneity on solid-state NMR pulse se- PROGRAM — BY DAY theory. quences are small and thus negligible for experimental implemen- TOPICS In nutation spectra, additional sidebands at multiples of the rotor tations. They could, however, play a more important role in the nu- PRIZE LECTURES frequency arise due to modulation of the rf phase. Significantly merical optimization of new pulse sequences. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] K. Aebischer et al. , Magn. Reson. Discuss. (preprint) 2021, https://doi.org/10.5194/mr-2021-43 [2] Z. Tošner et al., J. Magn. Reson. 2017, 284, 20-32. [3] P. Tekely, M. Goldman, J. AUTHOR INDEX Magn. Reson. 2001, 1, 135-141. IN022 PARALLEL SESSIONS / INVITED SPEAKER / BENCHTOP AND LOW-FIELD NMR 88 BERNHARD BLÜMICH PROGRESS IN COMPACT NMR Institut für Technische und Makromolekulare Chemie, RWTH The era of compact NMR started in the first half of the 1970ties with the advent of tabletop NMR instruments, most Aachen University, Aachen, notably the Bruker Minispec relaxometer and the Newport Analyzer, a relaxometer from a company sunsequently Germany acquired by Oxford Instruments [1]. In took about 50 years to figure out how to cope with the tolerances of permanent magnet blocks so that they can be assembled reliably to make small permanent magnets with field homoge- neity sufficiently good to resolve the 1H chemical shifts of small molecules in solution. Today multi-nuclear NMR spectroscopy with compact tabletop instruments is an established analytical method in many academic and indus- trial laboratories [2]. Arguably the miniaturization of compact permanent magnets benefitted from the advances in profiling the strayfield of inside-out NMR sensors used for logging oil wells and related compact strayfield relaxometers for nondestructive materials testing [3,4]. Recent progress in instrumentation and applications of compact NMR is reported. In particular, methods of map- ping the sensitive volume of strayfield-sensors like the NMR-MOUSE and compact magnets have been developed [5], the NMR-MOUSE has been employed in unconventional outdoor scenarios like Yellowstone National Park and the fire damaged library of the Glasgow School of Art [6], a compact and open permanent magnet has been built and passively shimmed for chemical shift resolved Overhauser DNP experiments [7], and a high-pressure setup has been constructed and tested for compositional analysis of gas mixtures at pressures of up to 200 bar [8]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] B. Blümich, J. Magn. Reson. 2019, 306, 27-35. [2] B. Blümich, K. Singh, Angew. Chem. Int. Ed. 2018, 57, 6996-7010. [3] J.A. Jackson, L.J. Burnett, INVITED AND PROMOTED LECTURES J.F. Harmon, J. Magn. Reson. 1980, 41, 411-421. [4] B. Blümich, J. Perlo, F. Casanova, Prog. Nucl. Magn. Reson. Spectr. 2008, 52, 197-269. [5] T. Überrück, C. Rehorn, R. Höhner, B. Blümich, J. Magn. Reson. 2018, 296, 169-175. [6] B. Blümich, D. Jaschtschuk, C. Rehorn, in: S. Haber-Pohlmeier, B. Blümich, L. Ciobanu, eds., POSTERS Magnetic Resonance Microscopy. Instrumentation and Applications in Engineering, Life Science and Energy Research, Wiley-VCH, Weinheim, 2021, in press. [7] T. ADVERTISERS Überrück, M. Adams, J. Granwehr B. Blümich, J. Magn. Reson. 2020, 314 , 10267. [8] A. Duchowny, P.M. Dupuy, H.C. Widerøe, O.J. Berg, A. Fanes, A. Paulsen, H. , AUTHOR INDEX Thern, O. Mohnke, B. Blümich, A. Adams, manuscript in preparation, 2021. IN023 PARALLEL SESSIONS / INVITED SPEAKER / BENCHTOP AND LOW-FIELD NMR 89 DMITRY BUDKER1,2,3 ZERO- TO ULTRALOW-FIELD (ZULF NMR): 1Institut für Physik, Johannes RECENT PROGRESS AND NEW DIRECTIONS Gutenberg Universität-Mainz, Mainz, Germany, 2GSI Helmholtzzentrum In this talk, we will recall how it is possible to do NMR without magnets, survey the recent progress, and present new für Schwerionenforschung, directions being pursued in our laboratory and elsewhere. Helmholtz-Institut Mainz, Mainz, Germany, 3University of California, Berkeley, California, USA INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PT030 PARALLEL SESSIONS / PROMOTED TALK / BENCHTOP AND LOW-FIELD NMR 90 JONATHAN FARJON 1, OPTIMIZING ENZYME- AND CELL-BASED BIOPROCESSES WITH DYLAN BOUILLAUD 1, MOBILE FLOW NMR ALPER SOYLER 2, MECIT OZTOP 2, Better optimizing biochemical processes induced by free enzymes 2- Several microalgae have the ability to produce lipids thanks to a OLIVIER GONÇALVES 3, or by cells, is crucial to improve the production of high added value metabolic redirection provoked by nitrogen starvation conditions. PATRICK GIRAUDEAU 1 commercial products in pharmaceutics, cosmetics, food and energy In the context of following bioprocesses by low field NMR [4], a 43 1Université de Nantes, CNRS, sciences. NMR is essential to the optimization and control of biopro- MHz apparatus was used for the first time for non-invasive moni- Nantes, France, 2 cesses since it provides both structural and quantitative information toring of entire microalgae in their cultivation medium. Thanks Department of Food Engineering, Middle East Technical University, in a non-destructive fashion. In this context, emerging benchtop to the implementation of WATERGATE W5 scheme [5], the water Ankara, Turkey, NMR instruments are particularly attractive since they are cheap, peak representing more than 95% of the culture could efficiently be 3Université de Nantes, GEPEA, St. mobile and compatible with industrial settings. However, at low- removed: 27,000 times less than in 1H. The main peak from in vivo Nazaire, France er fields, the limits of NMR are worsened: low sensitivity and weak lipids is then observable after 1h of W5 acquisition and can be as-spectral resolution are detrimental to the analytical performance. To signed and quantified as the total lipids [6]. To monitor lipid in-cell overpass such limitations, we implemented NMR techniques using accumulation, a benchtop spectrometer was coupled to a photo-pulse field gradients in diffusion NMR, pure-shift methods, ultrafast bioreactor, an automated device for microalgae cultivation. For the 2D NMR and schemes for removing the water signal in biological first time, the real-time online and in vivo access to lipid production samples, on a 1 T apparatus equipped with a gradient coil [1]. kinetics during 3 weeks by compact NMR was obtained with a very similar profile to GC offline reference method [7]. Our recent results 1- In the field of food processes, we showed that quantitative WET- with compact flow NMR monitoring open the avenue for improving 180-NOESY [1] is the most suitable tool able to selectively remove INTRODUCTION enzymatic and in vivo cell bioprocesses involving the real time de- the water signal to monitor under flow two key enzyme-catalyzed SPONSORS tection and quantification of valuable metabolites. bioprocesses in food industry: sucrose inversion for enhancing the PROGRAM sweetness [2] or for producing lactose free milk [3] for intolerant PROGRAM — BY DAY persons. TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] B. Gouilleux, J. Farjon, P. Giraudeau, J. Magn. Reson. 2020, 319, 106810-106822. [2] A. Soyler, D. Bouillaud, J. Farjon, P. Giraudeau, M. H. Oztop, LWT Food ScI. Tech. 2020, 118, TUTORIAL LECTURE 108832-108839. [3] A. Soyler, S. Cikrikci, D. Bouillaud, C. Cavdaroglu, J. Farjon, P. Giraudeau, M. H. Oztop, LWT Food ScI. Tech. 2021, 139, 110557-110567. [4] D. Bouillaud, J. Farjon, O. Gonçalves, P. Giraudeau, Magn. Reson. Chem. 2019, 57, 794-804. [5] X. Mao, L. Liu, H. Huang, J. Nicholson, J. Lindon, J. Magn. Reson. 1998, 132, 125-129. [6] D. Bouillaud, T. Cordier-Castaing, A. V. Heredia INVITED AND PROMOTED LECTURES Marquez, D. Drouin, O. Gonçalves, J. Farjon P. Giraudeau, Algal Res. 2019, 43, 101624-101630. [7] D. Bouillaud, D. Drouin, B. Charrier, C. Jacquemmoz, P. Giraudeau, J. Farjon, O. Gonçalves, , POSTERS Process Biochemistry 2020, 93, 63–68. ADVERTISERS Acknowledgements: The authors acknowledge support from the Region Pays de la Loire (“Pari Scientifique Régional AMER-METAL”), the French National Center for Scientific Research AUTHOR INDEX (“Osez l’Interdisciplinarité !” RMN-(ME)2-TAL). A. Soyler thanks the STSM program under COST action CA15209 EURALAX. IN024 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULES AND INTERACTIONS II 91 LUKÁŠ ŽÍDEK NMR REVEALS STRUCTURE-FUNCTION RELATIONSHIP IN Faculty of Science and CEITEC, DISORDERED PROTEINS: MAP2C AS AN EXAMPLE Masaryk University, Brno, Czech Republic It is well documented that intrinsically disordered proteins (IDPs) polymerase, with almost uniform preference of polyproline II con-play important biological functions, especially in signalling pro- formations [5]. Analysis of relaxation and paramagnetic relaxation cesses. However, it is less clear how is the biological activity of IDPs enhancement showed formation of a transient compact structure, related to their structure and dynamics. Favourable relaxation prop-not observed in Tau. erties of IDPs and development of non-uniform sampling methods Features related to the biological function of MAP2c were also stud- [1] make NMR an excellent tool for atomic-resolution studies of the ied by NMR. Line-broadening due to interaction with well-ordered structure-function relationship in large IDPs. Investigation of a mi-proteins allowed us to quickly identify interaction sites for several crotubule associated protein 2c (MAP2c) is presented as an example. binding partners. Overlap of the regions interacting with tubulin MAP2c is a splicing variant of the MAP2 gene expressed in devel- and with regulatory proteins (14-3-3, plectin) explains the mech- oping brain neurons. MAP2c resembles Tau in its ability to control anism of modulation of the regulatory activity of MAP2c by addi-dynamics of microtubules, but MAP2c and Tau differ in amino-acid tional protein-protein interactions [4]. Distinct amide frequencies sequences of their N-terminal domains and in cellular localization. of phosphorylated serine and threonine, and selective labelling of Compared to Tau and despite of its biological importance, MAP2c is tyrosine make measurement of phosphorylation kinetics possible much less studied than Tau. for all phosphorylated residues. Distinct phosphorylation patterns and kinetics of MAP2c and Tau represent another level of specific Almost complete resonance assignment of MAP2c, relying on 5D regulation of biological activity [6]. Furthermore, MAP2c, but not INTRODUCTION non-uniformly sampled experiments [2], made detailed studies of Tau, specifically binds regulatory subunit of an important protein SPONSORS MAP2c structure possible. The chemical shifts were converted to kinase, PKA. Positions of residues involved in the mentioned regu- PROGRAM populations of secondary structures by the ASTEROIDS analysis latory events strongly correlate with the identified local pre-struc- PROGRAM — BY DAY [3]. The results revealed numerous local pre-structured alpha-hel- tured motifs, and with the formation of the large compact structure. TOPICS ical or polyproline II motifs [4], in contrast to another IDP studied PRIZE LECTURES in the group, C-terminal domain of delta subunit of bacterial RNA PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] K. Kazimierczuk et al., J. Magn. Reson. 2008, 192, 123-130. 356-365. [2] J. Nováček et al., J. Biomol. NMR. 2013, 56, 291-302. [3] G. Nodet et al., J. Am. Chem. Soc. 2009, 131, 17908- ADVERTISERS 17918. [4] S. Jansen et al., J. Biol. Chem. 2017, 292, 6715-6727. [5] V. Kubáň et al., J. Am. Chem. Soc. 2019, 141, 16817-16828. [6] K. Melková et al., J. Biol. Chem. 2018, 293, 13297-13309. AUTHOR INDEX Acknowledgements: This work has been supported by the Czech Science Foundation, Grant GA20-12669S. IN025 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULES AND INTERACTIONS II 92 IRREM-LAAREB A FOUR-DOMAIN INTRAMOLECULAR FUZZY COMPLEX MOHAMMAD, IN C-SRC ESTEFANIA RUSCA, VICTOR RUIZ, The study of the myristoylated, intrinsically disordered region of c-Src, the first discovered oncogene, is a repre-ALEJANDRO sentative example of the paradigm shift towards understanding the integration of intrinsically disordered regions FERNANDEZ, and globular folded domains present in the majority of eucharyotic proteins implicated in high-order regulatory MIQUEL PONS processes. Bimolecular NMR group, Department of Inorganic and In this talk we shall present unpublished work on the use of NMR to characterize the entire 250-residue regulatory Organic Chemistry, University of region of c-Src, including the disordered SH4 and Unique domains, and the two globular regulatory domains SH3 Barcelona, Barcelona, Spain and SH2. This work extends our previous findings on the intramolecular fuzzy complex formed by the disordered regions and the SH3 domain [1, 2], and also on the modulation of c-Src membrane anchoring by competition between intra- and intermolecular interactions involving the myristoylated N-terminus of c-Src[3,4]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] M. Arbesú et. al. Structure. 2017, 25, 630-640, [2] M Arbesú, M Pons, Archives Biochem Biophys. 2019, 677, 108161. [3] A L Le Roux, et al. iScience. INVITED AND PROMOTED LECTURES 2019, 12, 194-203. [4] I L Mohammad, et al. BioMol Concepts, 2019, 10, 25-36. POSTERS Acknowledgements: This work was supported by grants BIO2016-78006-R; PID2019-104914RB-I00; and the Fundación Maria Francisca de Roviralta. We ADVERTISERS acknowledge the use of the Spanish “Instalación Científica Technologica Singular “ Red de Laboratorios de RMN de biomoléculas. The authors acknowledge AUTHOR INDEX the support and the use of resources of Instruct-ERIC internship application APPID 783. IN026 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULES AND INTERACTIONS II 93 LAURA MARIÑO PÉREZ1, CAPTURING THE HIGH-RESOLUTION STRUCTURE FRANCESCO S. IELASI2, OF A LOW-POPULATED AROMATIC RING FLIPPING LUIZA M. BESSA1, DAMIEN MAURIN1, INTERMEDIATE JAKA KRAGELJ1, MARTIN BLACKLEDGE1, Aromatic residues cluster in the core of folded proteins, where they stabilize the structure through multiple inter-NICOLA SALVI1, actions. NMR studies carried out in the 1970s surprisingly demonstrated that aromatic side chains can undergo ring GUILLAUME BOUVIGNIES3, flips, i.e. 180° rotations around the c dihedral angle, despite their central role in maintaining the protein fold [1-3]. 2 ANDRÉS PALENCIA2, It was suggested that large-scale breathing motions of the surrounding protein environment are necessary to ac- MALENE RINGKJØBING commodate these ring flipping events. However, the structural details of these motions were previously unknown. JENSEN1 Here, we uncover the structural rearrangements accompanying a ring flipping event of a buried tyrosine residue 1Institut de Biologie Structurale (IBS), in an SH3 domain. Using 1HN and 15N relaxation dispersion experiments [4-5], we show that the tyrosine side chain Grenoble France 2 flips to a low-populated (3%) excited state corresponding to a ring flipping intermediate. Based on a proteome-wide Institute for Advanced Biosciences (IAB), Grenoble, France sequence analysis, we design single point mutations that invert the relative populations of the ground and excited 3Ecole Normale Supérieure (ENS), Paris, state allowing us to capture the high-resolution structure of the ring flipping intermediate by X-ray crystallography. France Our results reveal how hydrogen bonds and CH-p interactions are rearranged in the intermediate providing unprecedented structural insight into the protein breathing motions associated with aromatic ring flipping. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] I. D. Campbell, C. M. Dobson, R. J. Williams, Proc. R. Soc. Lond. B, Biol. Sci. 1975, 189, 503-509. [2] K. Wüthrich, G. Wagner, FEBS Lett. 1975, 50, ADVERTISERS 265-268. [3] G. Wagner, A. DeMarco, K. Wüthrich, Biophys. Struct. Mech. 1976, 2, 139-158. [4] D. F. Hansen, P. Vallurupalli, L. E. Kay, J. Phys. Chem. B. 2008, 112, AUTHOR INDEX 5898-5904. [5] T. Yuwen, L. E. Kay, J. Biomol. NMR 2019, 73, 641-650. PT031 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULES AND INTERACTIONS II 94 GYULA BATTA1, DYNAMIC STRUCTURES AND STRESS INDUCED ANDRÁS CZAJLIK1, UNFOLDING OF SMALL ANTIMICROBIAL PROTEINS ÁDÁM FIZIL1, DOROTTYA HAJDU1, Small antimicrobial disulfide proteins like AFP, PAF, PAFC, PAFB, NFAP, NFAP2 are famous for their antifungal activÁGNES BATTA1, ity and some of them exhibit anti- Candida or even anti-corona virus activity[1]. They are harmless for mammalian ZOLTÁN GÁSPÁRI2, cells however, their diverse mode of action is not fully understood[2]. These miniproteins (50-60 aa) are produced LÁSZLÓ GALGÓCZY3, by filamentous fungi and they form β-barrel tertiary structures. Folded forms are stabilised by 3-4 disulfide bridg-FLORENTINE MARX4 es and therefore believed to exist as rock hard entities in spite of many charged residues (e.g. lysines). We have 1University of Debrecen, Structural shown[3] by cold & heat unfolding, 15N-relaxation, 15N chemical exchange saturation transfer (CEST) and ensemble Biology Research Group, Debrecen, molecular dynamics (MD) calculations that this is not the case. Even under the conditions of maximum protein sta-Hungary 2Faculty of Information Technology bility nuclear magnetic resonance (NMR) invisible protein states may persist due to conformational and dynamical and Bionics, Pázmány Péter Catholic heterogenity. The hidden nature of these states for everyday NMR methods may be explained by the low popula- University, Budapest, Hungary 3 tions and/or fading effects due to exchange between two or more states in the intermediate time scale (ms-us) re-Department of Biotechnology, Faculty of Science and Informatics, gime. Chemical stress induced unfolding is also interesting since in pharmaceutical research as co-solvent, DMSO University of Szeged, Szeged, Hungary is often used. Here we present DMSO induced unfolding of PAF and variants as followed by NMR and differential 4Institute of Molecular Biology, Biocenter, Innsbruck Medical scanning calorimetry (DSC), including thermodynamics parameters and structures. Partially unfolded states can be University, Innsbruck, Austria biologically relevant, e.g. connected to disulfide shuffling or S-S bond chirality switching. In general, less structured intermediates can be preferred for conformational-selection upon molecular recognition. In fact, there should not be a sharp boarder between the folded and intrinsically disordered proteins (IDPs) world and it is well known that INTRODUCTION many folded proteins have disordered regions. Practical consequences may have impact on the validation of MD SPONSORS simulations or protein concentration measurements. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] A. Huber, L. Galgóczy, G. Váradi, J. Holzknecht, A. Kakar, N. Malanovic, R. Leber, J. Koch, M. A. Keller, G. Batta, G. K. Tóth and F. Marx, Biochimica TUTORIAL LECTURE Et Biophysica Acta-Biomembranes 2020, 1862. [2] A. Czajlik, J. Holzknecht, L. Galgóczy, L. Tóth, P. Poór, A. Ördög, G. Váradi, A. Kuhbacher, A. Borics, G. K. Tóth, F. Marx and G. Batta, International Journal of Molecular Sciences 2021, 22. [3] Á. Fizil, Z. Gáspári, T. Barna, F. Marx and G. Batta, Chemistry-A European Journal INVITED AND PROMOTED LECTURES 2015, 21, 5136-5144. POSTERS Acknowledgements: The research was supported by the EU and co-financed by the European Regional Development Fund under the projects ADVERTISERS GINOP-2.3.2-15-2016-00008 and GINOP-2.3.3-15-2016-00004, and also by the Hungarian National Grant OTKA ANN 110 821, the Austrian Science Fund AUTHOR INDEX (I3132-B21) to F.M. L.G. was financed by the FK 134343 project of the Hungarian National Research, Development and Innovation (NKFIH) Office. PT032 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULES AND INTERACTIONS II 95 STASĖ BIELSKUTĖ1,2, HEAVY WATER ENHANCES THE LIQUID-LIQUID PHASE CARLA GARCIA-CABAU1,2, SEPARATION OF A SUB-DOMAIN OF THE ANDROGEN MARTA FRIGOLÉ-VIVAS1,2, ELZBIETA SZULC1,2, RECEPTOR EVA DE MOL1,2, MIREIA PESARRODONA1,2, Phase equilibria of intrinsically disordered proteins (IDPs) are sensitive to changes in solution conditions, such as JESÚS GARCÍA1,2 AND concentration of an IDP, ionic strength or temperature, and this can be used to study the driving forces in biomo-XAVIER SALVATELLA1,2,3 lecular condensation formation by liquid-liquid phase separation (LLPS) in vitro. Biomolecular condensates can 1 have important roles in many cellular functions and dysregulation of their assembly and disassembly can be linked Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute to various diseases, including cancer. In our work, we have studied the LLPS of the transcriptional activation unit of Science and Technology, Barcelona, (Tau-5*) of the activation domain (AD) of the androgen receptor (AR), a transcription factor that plays a role in the Spain 2 development of the male phenotype and is an atractive therapeutic target for prostate cancer and castration resis-Joint BSC-IRB Research Programme in Computational Biology, Barcelona, Spain tance prostate cancer, which currently is incurable [1]. In particular, we studied the effect that heavy water (D O), 2 3ICREA, Barcelona, Spain which is used in small amounts as a co-solvent in solution NMR to correct fluctuations in the magnetic field, has on the phase equilibrium of the protein. Tau-5*, as well as the full length intrinsically disordered AD [2], undergoes lower critical solution temperature (LCST) LLPS, which is driven at least in part by hydrophobic interactions. In our study, we show that even small fractions of H O replaced with D O decrease the cloud point of LLPS of Tau-5*, likely reflecting a stabilization of 2 2 the hydrophobic interactions that drive condensation. D O phenomenon to enhance hydrophobic interactions had 2 INTRODUCTION already been known from protein folding and aggregation studies, where 100 % H O is replaced by D O. Here we 2 2 SPONSORS show that even 10 % D O can reduce the cloud point of Tau-5* LLPS by 8 oC and 50 % by 20 oC, that represent a very 2 PROGRAM profound alteration of the phase diagram of an IDP. Therefore, it is important to take this effect into consideration PROGRAM — BY DAY when studying phase separation phenomena with biophysical methods that require using D O as a co-solvent, such 2 TOPICS as solution NMR or EPR. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] E. De Mol, R.B. Fenwick, C.T.W. Phang, V. Buzon, E. Szulc, A. de la Fuente, A. Escobedo, J. Garcia, C.W. Bertoncini, E. Estebanez-Perpina, I.J. INVITED AND PROMOTED LECTURES McEwan, A. Riera, X. Salvatella, ACS Chem. Biol. 2016, 11, 2499-2505. [2] J.J. Bouchard, J.H. Otero, D.C. Scott, E. Szulc, E.W. Martin, N. Sabri, D. Granata, M.R. POSTERS Marzahn, K. Lindorff-Larsen, X. Salvatella, B.A. Schulman, T. Mittag, Molecular Cell 2018, 72, 19-36. ADVERTISERS Acknowledgements: S.B. acknowledges a PhD fellowship awarded by IRB in the 2020 call and funding from AGAUR (2017 SGR 324), MINECO (BIO2015- AUTHOR INDEX 70092-R) and the European Research Council (CONCERT, contract number 648201). PT033 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULES AND INTERACTIONS II 96 PANAGIOTA S. GEORGOULIA1‡, PROTEIN PHOSPHORYLATION ASSIGNMENT BY FAST DMITRY M. LESOVOY2‡, TAMMO DIERCKS3, HIGH-RESOLUTION FOSY NMR. IRENA MATEČKO-BURMANN4,5, BJÖRN M. BURMANN1,4, EDUARD V. BOCHAROV2, Phosphorylation is a prototypical example of post-translational modifications (PTMs) that dynamically modulate WOLFGANG BERMEL6, VLADISLAV Y. protein function, where dysregulation is often implicated in disease. NMR provides information on the exact loca-OREKHOV1* tion and time course of PTMs with atomic resolution and under nearly physiological conditions, including inside 1Department of Chemistry and Molecular Biology, University of living cells, but requires unambiguous prior assignment of affected NMR signals to individual atoms. Yet, exist-Gothenburg, Gothenburg, Sweden 2 ing methods for this task base on a global, hence, costly and tedious NMR signal assignment that may often fail, Department of Structural Biology, Shemyakin-Ovchinnikov, Institute of Bioorganic Chemistry RAS, Moscow, Russia; Research especially for large intrinsically disordered proteins (IDPs). Here we introduce a suite of 2D Focused Spectrosco-Center for Molecular Mechanisms of Aging and Age-related pY (FOSY) experiments as a fast, sensitive and robust method to rapidly obtain only the relevant local NMR signal Diseases, Moscow; Institute of Physics and Technology (State assignment. By employing the long-overlooked concept of selective polarisation transfer (SPT)[2], FOSY focusses University), Dolgoprudny, Russia 3NMR Platform, CiC bioGUNE, Bld. 800, Parque Tecnológico de onto one coupled nuclear spin system (i.e. a residue affected by PTM) at a time by selecting three to five known fre-Bizkaia, Derio, Spain quencies and, thus, provides the spectral dispersion equivalent to a 6D-7D experiment in only two dimensions and 4Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg, Gothenburg, Sweden with an efficiency and versatility higher than achievable by traditional broadband experiments. We demonstrate 5Department of Psychiatry and Neurochemistry, University of the efficiency of FOSY by assigning, in just a few hours, two phosphorylation sites of proline-dependent glycogen Gothenburg, Gothenburg, Sweden 6 synthase kinase 3 beta (GSK3β) in human Tau40, an IDP of 441 residues. The new approach will benefit NMR studies Bruker BioSpin GmbH, Silberstreife, Rheinstetten, Germany of protein hotspots, as sites involved in molecular interactions and conformational changes as well as assignment bottlenecks. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] D.M. Lesovoy, P.S. Georgoulia, T. Diercks, I. Matečko-Burmann, B.M. Burmann, E.V. Bocharov, W. Bermel, & V.Y. Orekhov (2021), TUTORIAL LECTURE arXiv:2101.00605 [2] P. Pelupessy, E. Chiarparin, Concepts Magn. Reson. 2000, 12, 103-124; N. Khaneja, B. Luy, S.J. Glaser, PNAS 2003, 100, 13162-66; D.M. INVITED AND PROMOTED LECTURES Korzhnev, V.Y. Orekhov, L.E. Kay, J. Am. Chem. Soc. 2005, 127, 713-21; A.L. Hansen, E.N. Nikolova, A. Casiano-Negroni, H.M. Al-Hashimi , J. Am. Chem. Soc. 2009, 131, 3818-19; E. Walinda, D. Morimoto, M. Shirakawa, K. Sugase , J. Biomol. NMR 2017, 68 (1), 41-52. POSTERS ADVERTISERS *Correspondence: PSG p.s.georgoulia@gmail.com & VYO vladislav.orekhov@nmr.gu.se AUTHOR INDEX ‡These authors contributed equally IN027 PARALLEL SESSIONS / INVITED SEAKER / MRI IN MATERIAL SCIENCE AND BIOMOLECULAR APPLICATIONS 97 MELANIE M. BRITTON IN OPERANDO MAGNETIC RESONANCE IMAGING FOR School of Chemistry, University of NEXT-GENERATION BATTERY CHEMISTRIES. Birmingham, Birmingham, U.K. The demand for improved batteries is driving research towards the development of new electrode and electrolyte materials. However, the search for optimised materials can fall short in our aim of developing improved batteries. This is because the improvement of individual battery components cannot, on their own, provide the step-change in performance we are searching for. It is the collective performance of materials that is going to enable us to make high-performance batteries for the future. As battery components are typically a composite of materials, it is important to be able to distinguish the properties of the material from the assembled component. Also, as components work cooperatively, we need to develop analytical techniques that enable us to characterise the performance of the battery holistically. In the last 10 years, magnetic resonance imaging (MRI) has shown promise for non-invasively visualising the spatial distribution, speciation, and mobility of molecules and ions in batteries, across electrodes and electrolytes, integrating atomic information across mesoscopic and macroscopic length scales [1,2]. In this talk, I will present MRI of battery chemistry, under working conditions (in operando), and provide examples of 1H, 19F, 7Li and 23Na MRI for lithium-ion (LIBs), sodium-ion (NIBs) and Zn-air (ZABs) batteries [3,4]. In LIBs and NIBs, it is possible to image directly the electroactive species using 7Li or 23Na MRI. While 67Zn is NMR active, its properties do not lend themselves to MRI and so electroactive Zn species in ZABs cannot be observed directly. However, ZAB battery chemistry can be visualised using 1H MRI of the aqueous electrolyte, which is sensitive to the Zn-oxygen electrochemistry of the battery [3]. The use of 1H MRI is also valuable in LIBs and NIBs, enabling the vi- INTRODUCTION sualisation of microstructural changes and dendrite formation with greater spatial and temporal resolution than by SPONSORS 7Li or 23Na MRI. Operando 1H, 7Li and 23Na MRI experiments are presented, which identify and map electroactive PROGRAM species across the electrode and electrolyte during charge cycling and galvanostatic plating. In the case of NIBs, the PROGRAM — BY DAY formation and evolution of dendrites are observed and mapped by 23Na MRI and their 3D microstructure visualised TOPICS by 1H MRI of the electrolyte [4]. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] M.M. Britton, Prog. Nucl. Magn. Reson. Spec. 2017, 101, 51-70. [2] M.M. Britton, Chemphyschem 2014, 15, 1731-1736. [3] M.M. Britton et al. J. Phys. AUTHOR INDEX Chem. Lett. 2013, 4, 3019-3023. [4] J. M. Bray et al. Nat. Commun. 2020, 11, 2083. IN028 PARALLEL SESSIONS / INVITED SPEAKER / MRI IN MATERIAL SCIENCE AND BIOMOLECULAR APPLICATIONS 98 WŁADYSŁAW P. WĘGLARZ NOT ONLY MICE AND RATS - ZTE MR IMAGING OF Institute of Nuclear Physics Polish ROCKS AT 9.4T Academy of Sciences, Kraków, Poland Magnetic Resonance Imaging at the high magnetic field is successfully used for biomedical work, especially in in-vi-vo conditions, thanks to superior signal to noise ratio (SNR) and availability of a multitude of advanced imaging techniques. Most of these techniques are however useless or of limited use for the research of porous rocks, due to magnetic susceptibility-induced artifacts, compromising the observed NMR signal at high fields. Because of that the nuclear magnetic resonance and imaging performed at a low magnetic field are typically chosen as a useful tool for the laboratory-scale petrophysical characterization of oil-bearing rocks, as it assures the minimization of the influence of magnetic-susceptibility-induced internal gradients. The typical choice of the pulse sequence for MRI of such samples are Single Point Imaging (SPI), or its variants (e.g. SPRITE), which however have high demand on imaging gradient and/or limitations for the achievable 3D resolution in a reasonable experimental time [1-2]. Application of the zero echo time (ZTE) pulse sequence improves the possibility for time effective 3D imaging of samples with short T [3-5]. Collection of the set of data points on FID during the TR period significantly accelerates 2 measurements for 3D imaging, comparing to SPI. ZTE is a purely frequency-encoded method, and acquiring of the signal starts immediately after excitation using, e.g. , radial centre-out trajectories, without the need for preceding encoding gradients. In the present lecture the application of 3D ZTE imaging for assessment of local hydration level in highly porous INTRODUCTION rocks (dolomite), as well as tight sandstone rocks are presented [4,5]. Correlation of the obtained results of the water SPONSORS content and spatial distribution in rock material, with the low field MR relaxometry as well as with gravimetric data PROGRAM provides interesting insight into the interpretation of the relaxation data in terms of assessment of sample’s poros- PROGRAM — BY DAY ity. Correlation with high-resolution micro-CT images of the same rock samples allows for direct assessment of the TOPICS role of the sample’s composition and structure in the dynamics of water hydration into the porous rock’s structure. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] S. Emid, J.H.N. Creyghton, Physica B & C 1985, 128 (1), 81-83. [2] S.D. Beyea, B.J. Balcom, P.J. Prado, A.R. Cross, C.B. Kennedy, R.L. Armstrong, INVITED AND PROMOTED LECTURES T.W. Bremner, J. Magn. Reson. 1998, 135 (1), 156-164. [3] M. Weiger, K.P. Pruessmann, eMagRes, 2012, 1:311-322. [4] Węglarz WP, Krzyżak A, Stefaniuk M, Magn. POSTERS Reson. Imag. 2016, 34, 492-495,. [5] W.P. Węglarz, A. Krzyżak, G. Machowski, M. Stefaniuk, Magn. Reson. Imag., 2018, 47, 54-59. ADVERTISERS Acknowledgements: This work was partially supported by the research projects Blue Gas (BG2/ShaleCarp/14) and Applied Research Program (PBS2/A2/16/ - AUTHOR INDEX NMR-Rocks), financed by the National Centre for Research and Development, Poland. PT034 PARALLEL SESSIONS / PROMOTED TALK / MRI IN MATERIAL SCIENCE AND BIOMOLECULAR APPLICATIONS 99 WOUTER M. J. FRANSSEN1, UNRAVELLING MULTI-MODAL TRENDS IN FRANK J. VERGELDT1, QUANTITATIVE MRI DATA BY PHASOR ANALYSIS JOHN P. M. VAN DUYNHOVEN1,2, HERBERT VAN AMERONGEN1,3, Quantitative Magnetic Resonance Imaging (qMRI) methods are used extensively for non-invasive diagnostics in CAMILLA TERENZI1 medicine and rely on obtaining high contrast images, which are suitable for analysis to recognize diseased tissues, 1Laboratory of Biophysics, Wageningen based on either relaxation (T , T ) or diffusion processes. 1 2 University & Research, Wageningen, The Netherlands. Often, these qMRI data sets show exponential or Gaussian decays, which need to be fitted, per image voxel, to ex-2Unilever Food Innovation Centre, Wageningen, tract the characteristic relaxation lifetimes or diffusion coefficients. When these signal decays are multi-modal, The Netherlands. 3MicroSpectroscopy Centre, Wageningen they become hard to analyse and, for every image voxel, an a priori assumption must be made regarding the num- University & Research, Wageningen, The ber of parameters to be used for fitting. Netherlands. To circumvent this issue, phasor analysis, a known non-fitting method for the study of lifetime fluorescence data, has recently been introduced for qMRI data processing by our group [1]. In phasor analysis, a plot is made that maps the lifetime information simultaneously for all pixels within the images. Creating this plot requires no assumptions on the type or multi-modal character of the decay underlying the data. By exploiting inter-pixel correlation in phasor plots, unique information can be obtained, for instance about partial volume effects, that are not supplied by per-voxel fitting analyses. We demonstrate the use of phasor analysis for qMRI data, with applications ranging from medicine to porous me- dia. Moreover, we introduce a method to greatly improve the accuracy of phasor processing, in a way that turns out INTRODUCTION to be unique for qMRI data over the original application of phasor in lifetime fluorescence [2]. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] F. J. Vergeldt, A. Prusova, F. Fereidouni, H. van Amerongen, H. van As, T. W. J. Scheenen, A. N. Bader, Sci. Rep. 2017, 7, 861. [2] W. M. J. Franssen, POSTERS F. J. Vergeldt, A. N. Bader, H. van Amerongen, C. Terenzi, J. Phys. Chem. Lett. 2020, 11, 9152-9158. ADVERTISERS Acknowledgements: This work is part of the 4TU Precision Medicine program supported by High Tech for a Sustainable Future, a framework commissioned AUTHOR INDEX by the four Universities of Technology of The Netherlands (https://www.4tu.nl/en/news/!/393/awarding_hightech/). PT035 PARALLEL SESSIONS / PROMOTED TALK / MRI IN MATERIAL SCIENCE AND BIOMOLECULAR APPLICATIONS 100 ENZA DI GREGORIO1, SUPRAMOLECULAR ADDUCTS BETWEEN MACROCYCLIC LUCIANO LATTUADA2, GD-COMPLEXES AND POLYAROMATIC SYSTEMS: ALESSANDRO MAIOCCHI2, SILVIO AIME1, A ROUTE TO ENHANCE THE RELAXIVITY THROUGH GIUSEPPE FERRAUTO1, THE FORMATION OF HYDROPHOBIC INTERACTIONS. ELIANA GIANOLIO1 1University of Turin, Department of MRI is the election imaging technique for the diagnosis and mon- 1H-NMR of YbHPDO3A complex w or w/o of pyrene derivatives (14 Molecular Biotechnologies and Health itoring of numerous diseases. About 40–45% of MRI scans (ca. 38 T). Finally, the in vivo proof of concept of the enhancement of con-Sciences, Torino, Italy, 2Bracco Imaging Spa, Bracco Research million per year) are performed with the use of Gadolinium based trast was obtained by MRI of tumor-bearing mice pre and post injec-Centre, Colleretto Giacosa (TO), Italy. contrast agents (GBCAs). The recent findings related to NSF and tion of GBCA (7T) upon injection of Gd-HPDO3A (0.15 mmol/Kg) or Gd-retention strongly required caution in the use of GBCAs.1 Chem-Gd-HPDO3A/HPTS adduct (0.15 and 0.45 mmol/Kg). istry becomes central in looking for i) more stable and ii) more effi- A high binding affinity of macrocyclic GBCAs toward pyrene deriv- cient GBCAs (i.e. enhanced relaxivity). Different routes to enhance atives was observed. The supramolecular adducts display a signifi-relaxivity were exploited as i) the set-up of non-covalent binding in- cant increase of relaxivity. No enhancement was observed for linear teractions with macromolecules present in solution (e.g. albumin), GBCAs. This is due to the increase of the molecular reorientation ii) the increase of the number of coordinated or second sphere water time (tR) and second sphere water molecules (for the presence of molecules, iii) the increasing of prototropic exchange rates.2,3Herein, SO - and OH). we describe the increase of relaxivity attainable through reversible 3 binding interactions between the hydrophobic region of macrocy- NMR spectra of the Yb-HPDO3A/ pyrene mixture support the forma- INTRODUCTION clic GBCAs and SO3-/OH containing pyrene derivatives. tion of the supramolecular adduct. When HPTS/Gd-HPDO3A ratio SPONSORS is 3:1 (m/m), >90% of Gd-HPDO3A is in the associated adduct and Macrocyclic (ProHance, Gadovist, Dotarem) and linear (Magnevist, PROGRAM there is a 40% relaxation enhancement in respect to the value ob- Omniscan, MultiHance) GBCAs were tested. The increase of relax- PROGRAM — BY DAY served for Gd-HPDO3A alone (i.e. 6.5 mM-1s-1 vs. 9.2 mM-1s-1 in se- ivity upon the addition of SO -/OH containing pyrene derivatives 3 TOPICS rum). In T1w-MRI of tumor-bearing mice there is the increase of sig- was assessed by 1H-relaxometry and 1H- / 17O-NMR. The binding pa- PRIZE LECTURES nal enhancement from 53% (upon i.v. of only Gd-HPDO3A) to 125% rameters K (association constant) and R (relaxivity of the adduct) a b PLENARY LECTURES (upon i.v. of Gd-HPDO3A/HPTS adduct). By concluding, the reported between GBCAs and the pyrene derivatives were calculated by us- TUTORIAL LECTURE results show a novel tool to enhance the relaxivity of GBCAs through ing the PRE technique (0.5T). 1H NMRD profiles were measured w or INVITED AND PROMOTED LECTURES the formation of supramolecular adducts at clinical doses. w/o of pyrene derivatives at variable B0 (0.00024 to 1.5 T). Insights POSTERS into the formation of the adduct were obtained by high resolution ADVERTISERS AUTHOR INDEX References: [1] Aime S, et al. Magn Reson Imaging. . 2009, 30, 1259. [2] Caravan P. et al. Chem. Soc. Rev. 2006, 35, 512. [3] Caravan P. et al. Acc. Chem. Res.. 2009, 42, 851. PT036 PARALLEL SESSIONS / PROMOTED TALK / MRI IN MATERIAL SCIENCE AND BIOMOLECULAR APPLICATIONS 101 KAJA TUŠAR, MAGNETIC RESONANCE IMAGING WITH IGOR SERŠA NONLINEAR GRADIENT COILS Jožef Stefan Institute, Ljubljana, Slovenia Gradient coils are usually designed such that they produce spatially coils were connected to the gradient amplifiers of our system for linearly dependent magnetic fields along the direction of the static magnetic resonance microscopy based on a 2.35 T superconducting magnetic field. Specifically, three different gradient coils are used, magnet. Imaging with the nonlinear gradient coils was tested using each having a constant gradient of the magnetic field along one of the 2D spin-echo imaging sequence (without slice selection). One the three orthogonal spatial coordinates [1]. This approach results nonlinear gradient coil was used as the readout gradient and the in a simple relation between the frequency and the coordinate, other as the phase gradient. For the testing purposes was designed where one is proportional to the other. This relation makes possible also a special checkerboard patterned sample in a form of a 26 mm the introduction of the k-space, i.e., reciprocal space to the r-space, diameter disc of which 2 mm deep square holes were filled with gel. and also simple MR image reconstruction from the k-space data by Acquired signals were reconstructed in a standard manner using 2D multidimensional Fourier transformation. However, conventional Fourier transform. Since the signals were not k-space data, the ob-linear gradient coils have also some drawbacks, such as complex de- tained image was heavily distorted [2]. This was due to the nonlinear sign that usually leads to a higher inductance of the coils and there- relation between the frequency and spatial coordinates; for the test- fore more difficult fast gradient pulse switching. In this study we ed nonlinear gradient coils the frequency was inversely proportional examine the possibility of using nonlinear gradient coils for MRI. to the spatial coordinate (measured as the distance from the effective loop segment). The obtained image can also be interpreted as the 2D We designed a simple set of two orthogonal nonlinear gradient coils. spectrum and since the relation between the frequency and spatial Each of the coils was made of a single 114 mm by 40 mm rectangu- coordinates was known, the spectrum was transformed to the coor- INTRODUCTION lar loop of which two longer segments were parallel with the static dinate system where axes were spatial coordinates. In this coordinate SPONSORS magnetic field (nonessential for imaging) and two shorter ones were system the image of the sample was undistorted. PROGRAM perpendicular to it. One of the shorter segments lied in the imaging PROGRAM — BY DAY plane and was therefore producing considerable magnetic fields Results of the study confirmed that undistorted MR imaging with TOPICS along the direction of the static magnetic field, while the opposite nonlinear gradient coils is feasible. Since such coils have consider- PRIZE LECTURES segment was too far from the sample to have any significant effect ably less constrains in their design, this approach also enables opti- PLENARY LECTURES on imaging. Each of the loops was made of 50 turns of cooper wire mization of gradient coils for fast gradient pulse switching, which is TUTORIAL LECTURE and was tested with the maximum current of 3 A so that effective essential for several MR imaging methods and other applications, e.g. INVITED AND PROMOTED LECTURES current along the segments was up to 150 A. The nonlinear gradient diffusion measurements using PGSE, OGSE or MGSE sequences [3]. POSTERS ADVERTISERS References: [1] S. Hidalgo-Tobon, Conc. Magn. Res. A, 2010, 36A, 223. [2] S. Langlois, M. Desvignes, J. M. Constans, M. Revenu, J. Magn. Res. Imaging. 1999, 9, 821. [3] P. Callaghan, J. Stepisnik, AUTHOR INDEX Phys. Rev. Lett. 1996, 75, 4532. IN029 PARALLEL SESSIONS / INVITED SPEAKER / INTEGRATED STRUCTURAL BIOLOGY 102 TOBIAS MADL1,2 INTEGRATION OF NMR, SAXS, AND COMPLEMENTARY 1Gottfried Schatz Research TECHNIQUES FOR STRUCTURE DETERMINATION OF Center for Cell Signaling, Metabolism and Aging, BIOMOLECULAR COMPLEXES Molecular Biology and Biochemistry, Medical University of Graz, Graz, Austria Structural analysis of biomolecules is a key challenge in current biology and a prerequisite for understanding the 2BioTechMed-Graz, Graz, Austria molecular basis of essential cellular processes. The use of solution techniques is important for characterizing structure, complex formation and dynamics of biomolecules and biomolecular complexes. As experimental data for large biomolecules and biomolecular complexes are often sparse, it is advantageous to combine these data with additional information from other solution techniques. In my presentation I will show our recent achievements in integrating NMR data with complementary data from Small-Angle X-ray Scattering (SAXS), X-ray crystallography, electron microscopy, and mass spectrometry to study structure and dynamics of large proteins and biomolecular complexes [1-10]. By using our integrated approach we were able to provide a comprehensive and accurate description of protein complex structure and dynamics in a native-like environment. This underscores the central role of NMR for structure determination of biomolecular complexes and ensures its unique role and contributions in integrated structural biology approaches in the future. INTRODUCTION References: [1] Duchardt-Ferner, E., Juen, M., Bourgeois, B., Madl, T., Kreutz, C., Ohlenschlager, O. & Wohnert, J., Nucleic Acids Res. 2020, 48, 949-961. [2] SPONSORS Schopf, F. H., Huber, E. M., Dodt, C., Lopez, A., Biebl, M. M., Rutz, D. A., Muhlhofer, M., Richter, G., Madl, T., Sattler, M., Groll, M. & Buchner, J. Mol Cell, 2019, 74, 73-87 e8. [3] Rutz, D. A., Luo, Q., Freiburger, L., Madl, T., Kaila, V. R. I., Sattler, M. & Buchner, J., Nat Commun. , 2018, 9, 1472. [4] Zierer, B. K., Rubbelke, M., PROGRAM Tippel, F., Madl, T., Schopf, F. H., Rutz, D. A., Richter, K., Sattler, M. & Buchner, J., Nat Struct Mol Biol. 2016, 23, 1020-1028. [5] Rohl, A., Wengler, D., Madl, T., PROGRAM — BY DAY Lagleder, S., Tippel, F., Herrmann, M., Hendrix, J., Richter, K., Hack, G., Schmid, A. B., Kessler, H., Lamb, D. C. & Buchner, J., Nat Commun. 2015, 6, 6655. [6] TOPICS Rohl, A., Tippel, F., Bender, E., Schmid, A. B., Richter, K., Madl, T. & Buchner, J., EMBO Rep. 2015, 16, 240-9. [7] Gersch, M., Famulla, K., Dahmen, M., Gobl, C., Malik, I., Richter, K., Korotkov, V. S., Sass, P., Rubsamen-Schaeff, H., Madl, T., Brotz-Oesterhelt, H. & Sieber, S. A., Nat Commun. 2015, 6, 6320. [8] Lorenz, O. R., PRIZE LECTURES Freiburger, L., Rutz, D. A., Krause, M., Zierer, B. K., Alvira, S., Cuellar, J., Valpuesta, J. M., Madl, T., Sattler, M. & Buchner, J. Mol Cell. 2014, 53, 941-53. [9] Karagoz, PLENARY LECTURES G. E., Duarte, A. M., Akoury, E., Ippel, H., Biernat, J., Moran Luengo, T., Radli, M., Didenko, T., Nordhues, B. A., Veprintsev, D. B., Dickey, C. A., Mandelkow, E., TUTORIAL LECTURE Zweckstetter, M., Boelens, R., Madl, T. & Rudiger, S. G., Cell 2014, 156, 963-74. [10] Muller, R., Grawert, M. A., Kern, T., Madl, T., Peschek, J., Sattler, M., Groll, M. & Buchner, J., Proc Natl Acad Sci U S A. 2013, 110, 10183-8. INVITED AND PROMOTED LECTURES Acknowledgements: The work was supported by Austrian Science Fund (FWF) grants P28854, I3792, doc.funds BioMolStruct DOC 130 and DK-MCD W1226, POSTERS BioTechMed-Graz (Flagship project DYNIMO), the Austrian Research Promotion Agency (FFG) Grants 864690 and 870454; the Integrative Metabolism ADVERTISERS Research Center Graz; Austrian Infrastructure Program 2016/2017, and the Styrian Government (Zukunftsfonds). F.Z. was trained within the frame of the PhD AUTHOR INDEX program Molecular Medicine, Medical University of Graz. IN030 PARALLEL SESSIONS / INVITED SPEAKER / INTEGRATED STRUCTURAL BIOLOGY 103 PAU BERNADÓ STRUCTURAL BASES OF HUNTINGTON’S DISEASE Centre de Biochimie Structurale PATHOLOGICAL THRESHOLD. AN INTEGRATIVE (CBS), INSERM, CNRS, Université de Montpellier, Montpellier, STRUCTURAL BIOLOGY APPROACH France. Huntington’s disease (HD) is one of nine hereditary neurodegenerative disorders caused by an expansion of CAG triplet repeats beyond a pathological threshold. For HD, this expansion is located in the first exon of the huntingtin gene and results in an abnormally long poly-glutamine (poly-Q) tract within the N-terminus of the huntingtin protein (httex1). When the number of consecutive glutamines exceeds 35 (pathological threshold), the resulting mu- tant protein forms large cytoplasmic and nuclear aggregates, a hallmark of HD, and causes neuronal degeneration, especially affecting the neurons of the striatum. Aggregation, disease risk and age of onset correlate with the length of the poly-Q homo-repeat. The origin of a pathological threshold in HD and the other poly-Q related diseases remains poorly understood and different toxicity models for httex1 have been proposed. The highly repetitive nature of httex1 sequence, with long tracts of glutamines and prolines, hampers high-resolution investigations in solution by Nuclear Magnetic Resonance (NMR). Our group has developed a novel chemical biology approach that allows for the first time to obtain atomic-resolution information of httex1 independently of the poly-Q length [1]. Using this strategy we have derived structural models of a pathogenic and a non-pathogenic versions of httex1 with 46 and 16 consecutive glutamines, respectively [2]. The comparison of these models shows that in both cases the protein consists of an equilibrium of INTRODUCTION helical conformations involving different fractions of the poly-Q. We also show that both poly-Q flanking regions SPONSORS define the structure of the homo-repeat. Using Small-Angle Scattering and computational approaches we have PROGRAM shown the presence of long a-helices, suggesting a mechanism of toxicity that involves the formation of oligomeric PROGRAM — BY DAY species through coiled-coil interactions. Taken together, these observations provide the structural bases to under- TOPICS stand previous biophysical and functional data on httex1. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] A. Urbanek, A. Morató, F. Allemand, E. Delaforge, A. Fournet, M. Popovic, S. Delbecq, N. Sibille, P. Bernadó Angew. Chem. Int. Ed. Engl. 2018, INVITED AND PROMOTED LECTURES 57, 3598-3601. [2] A. Urbanek, M. Popovic, A. Morató, A. Estaña, C.A. Elena-Real, P. Mier, A. Fournet, F. Allemand, S. Delbecq, M.A. Andrade-Navarro, J. Cortés, POSTERS N. Sibille, P. Bernadó Structure. 2020, 28, 733-746. ADVERTISERS Acknowledgements: This work was supported by the European Research Council under the European Union’s H2020 Framework Programme (2014-2020) / AUTHOR INDEX ERC Grant agreement n° [648030]. PT037 PARALLEL SESSIONS / PROMOTED TALK / INTEGRATED STRUCTURAL BIOLOGY 104 TÖRNER RICARDA1; INTEGRATION OF NMR, KINETIC ASSAYS AND MICROSCOPY TO KUPREICHYK TATSIANA2; INVESTIGATE INHIBITION OF AMYLOID FIBRIL FORMATION BY GREMER LOTHAR2; AWAD RIDA1; CO-CHAPERONIN PREFOLDIN DAPHNA FENEL1; GANS PIERRE1; Dysfunction of proteostasis leads to the accumulation of misfold- We have established that PFD inhibits the formation of IAPP am- GUY SCHOEHN1; ed protein aggregates, as observed in many age-related amyloid yloid fibrils at sub-stoichiometric ratios. By using an integrative DIETER WILLBOLD2; diseases [1]. Among them is type II diabetes, caused by aggregation approach, we reveal the action mode of prefoldin during the fibril HOYER WOLFGANG2; of Islet amyloid polypeptide (IAPP) in pancreatic beta cells [2]. Nor- elongation process, responsible for this strong inhibitory effect. BOISBOUVIER JEROME1 mally, the quality control machinery, notably the chaperone net- Analysis of ThioflavinT- fibrillation assays show that PFD inhibits 1 work, keeps aggregation in check, but in cases of misfolding diseas- the secondary nucleation and elongation of IAPP fibrils, which was Univ. Grenoble Alpes, CNRS, CEA, Institut de Biologie Structurale (IBS), es these systems are incapacitated, either due to age or overwhelm further confirmed by an EM investigation of PFD fibril binding. To Grenoble, France. by more aggregation-prone protein species. So pharmacological enable NMR studies, assignment of the heterohexameric chapero-2Institut für Physikalische Biologie, targeting of the chaperone network is a promising avenue for find- nin PFD (90kDa) was achieved with the aid of an in-house developed Heinrich-Heine-Universität Düsseldorf, Düsseldorf, Germany ing disease altering therapeutics, requiring however a detailed un- isotope-aided NMR method [5]. Liquid state NMR interaction stud- derstanding of chaperone action. ies with inovative PRE-labeling on IAPP were used to investigate binding of PFD to IAPP and extract kinetic parameters. We demon- Here we report the mechanistic study of IAPP fibrillation inhibition strate that client binding takes place via two binding regions on by the cytosolic Hsp60 co-chaperonin prefoldin (PFD) [3], which has IAPP, which bind in a highly dynamic fashion inside the PFD cavity. previously been shown to interact with other amyloidogenic sub- INTRODUCTION strates [4]. Our strategy, based on integration of kinetic data and Our approach gives structural and mechanistic insights into the SPONSORS structural studies by microscopy and NMR spectroscopy, allows to process of chaperone inhibition of fibrillation, a first step towards PROGRAM elucidate the different inhibition pathways and provides structural modulation of the interaction for disease-prevention. PROGRAM — BY DAY insight on the interaction. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] J. Labbadia and R.J. Morimoto, Annu Rev Biochem. 2017, 84 435-464; [2] K. Pillay and P. Govender, BioMed research international 2013; [3] I. E. Vainberg et al., Cell 1998, 93, AUTHOR INDEX 863–73; [4] K. Sörgjerd et al., Biochemistry 2013,20 3532-3542; [5] R. Törner et al. , J. Biomol NMR 2020, 74, 83–94. PT038 PARALLEL SESSIONS / PROMOTED TALK / INTEGRATED STRUCTURAL BIOLOGY 105 RAPHAEL STOLL NMR-BASED STRUCTURAL INSIGHTS INTO Faculty of Chemistry and PHOTOSYSTEM II ASSEMBLY Biochemistry, Ruhr University of Bochum, Bochum, Germany Biogenesis of photosystem II (PSII), nature’s water-splitting catalyst, is assisted by auxiliary proteins that form transient complexes with PSII components to facilitate stepwise assembly events. Using cryo-electron microscopy, the structure of such a PSII assembly intermediate from Thermosynechococcus elongatus could be solved at 2.94 Å resolution [1]. It contains three assembly factors (Psb27, Psb28 and Psb34) and provides detailed insights into their molecular function [2]. Binding of Psb28 induces large conformational changes at the PSII acceptor side, which distort the binding pocket of the mobile quinone (QB) and replace the bicarbonate ligand of non-haem iron with glutamate, a structural motif found in reaction centres of non-oxygenic photosynthetic bacteria. NMR spectrosco-py reveals that the carboxy-terminus of Psb28 is rigidified upon CP47 binding due to creation of an intermolecular β-sheet (Fig. 1). Together, these results bring to light mechanisms that protect PSII from damage during biogenesis until water splitting is activated. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] Zabret et al. , Nature Plants 2021, 7, 524-538. [2] Cormann et al. , Biochemistry 2009, 48, 8768-8770. PT039 PARALLEL SESSIONS / PROMOTED TALK / INTEGRATED STRUCTURAL BIOLOGY 106 NICOLAS COUDEVYLLE1, NMR INSIGHT INTO THE RECRUITMENT OF ATG9 BY BARTŁOMIEJ BANAŚ2, ATG11 IN YEAST SELECTIVE AUTOPHAGY VERENA BAUMANN1, MARTINA SCHUSCHNIG1, Autophagy is a conserved catabolic pathway that mediates the degradation of substances within lytic compart- ANNA ZAWADZKA- ments. The degradation of these substances (from protein aggregates to damaged organelles), referred to as cargo, KAZIMIERCZUK2, WIKTOR is achieved through their sequestration within de novo formed double membrane organelles called autophago- KOŹMIŃSKI2 AND somes. In selective autophagy a specific cargo material is targeted for degradation. Selective autophagy starts with SASCHA MARTENS1. cargo recognition by a receptor and a scaffold protein (Atg11 in yeast, FIP200 for humans), which in turn triggers the 1Max Perutz Laboratories, University of recruitment of the autophagy machinery driving autophagosome formation around the cargo. Vienna, Vienna, Austria 2Biological and Chemical Research Vesicles containing the Atg9 protein are fascinating cogs of the autophagy machinery. These vesicles are 60 nm in Centre, Faculty of Chemistry, University diameter and contain 24 to 32 copies of the protein Atg9 (115 kDa) [1]. We have recently shown that Atg9 vesicles of Warsaw, Warsaw, Poland form platforms for the recruitment of the autophagy machinery as well as a membrane seed for phagophore expan- sion [2]. In yeast selective autophagy, the recruitment of Atg9 vesicles is mediated by the scaffold protein Atg11 (135 kDa) that interacts with the N-terminal domain of Atg9 [3]. In order to characterise the interaction between Atg11 and Atg9, we cloned, expressed and purified the N-terminal domain of Atg9 (residues 1 to 285). ITC titration showed that Atg9-NTD binds to Atg11 with a low micromolar affinity while the 1H-15N HSQC spectrum revealed that Atg9-NTD is largely disordered. Using 1H-15N HSQC based titration we were able to identify 2 Pro-Leu-Phe motifs in Atg9 that are mediating the binding to Atg11. Mutating these motifs to INTRODUCTION Pro-Ala-Ala completely abolished Atg11 binding in vitro as well as selective autophagy pathways in vivo. SPONSORS Our data provide structural insights into the first steps of phagophore formation. Additionally, we illustrated how PROGRAM NMR data driven mutagenesis can lead to a better understanding of in vivo processes. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Yamamoto, H., et al., J. Cell Biol., 2012, 198(2): p. 219-33. [2] Sawa-Makarska, J. et al. , Science, 2020, 369(6508). [3] He, C., et al. , J. Cell Biol. 2006, AUTHOR INDEX 175(6): p. 925-35. IN031 PARALLEL SESSIONS / INVITED SPEAKER / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 107 ANTONIN SOJKA, DEVELOPMENT OF A HIGH-FREQUENCY RAPID SCAN MATUŠ ŠEDIVÝ, ELECTRON SPIN RESONANCE SPECTROMETER ARTUR SOLODOVNIK, ADAM LAGIN, We report on the recent development of a high-frequency rapid scan electron spin resonance (FRASCAN) spec- TOMÁŠ LÁZNIČKA, trometer at the Brno University of Technology. The basic principle of frequency rapid scan will be explained and VINICIUS SANTANA, compared to conventional methods. The FRASCAN operates in induction mode using quasi-optics with a super- ANDRIY MARKO, heterodyne detection scheme. Fast frequency sweeps of the order of 1000 THz/s allows to access spin relaxation OLEKSII LAGUTA, of the order of 1 ns [1], in a frequency range of 80 GHz to 1100 GHz [2], at temperatures from 1.8 K to 300 K, and at PETR NEUGEBAUER magnetic fields up to 16 T. We developed several sample holders for performing measurements on liquids, oriented Central European Institute of single crystals, and air-sensitive samples, including the possibility of photo-excitation [3]. In addition, we developed Technology, Brno University of a carousel sample holder for pressed powders that accommodates up to 6 samples, avoiding the time-consuming Technology, Brno, Czech Republic event of loading the probe into the cryostat and cooling down process. The carousel holder can be used for quantitative ESR. The FRASCAN is controlled by a home-written software in LabView, allowing to run experiments in an automatic mode controlled by scripts. Frequency rapid scan experiments on an oriented single crystal of LiPc will be presented along with simulation for calculation of the relaxation times. Furthermore, additional capabilities of FRASCAN are demonstrated using frequency-detected magnetic resonance spectra as a function of the orientation for a single-crystal of copper acetate and frequency-field ESR maps for Mn and TEMPO. 12 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] O. Laguta, M. Tuček, J. van Slageren, P. Neugebauer, J. Magn. Reson. 2018, 296, 138–142. [2] P. Neugebauer, D. Bloos, R. Marx, P. Lutz, M. Kern, D. ADVERTISERS Aguila, J. Vaverka, O. Laguta, C. Dietrich, R. Clérac and J. van Slageren, Phys.Chem.Chem.Phys., 2018, 20, 15528. [3] A. Sojka, M. Šedivý, A. Solodovnik, A. Lagin, AUTHOR INDEX T. Láznička, V. Santana, A. Marko, O. Laguta and P. Neugebauer, to be published. IN032 PARALLEL SESSIONS / INVITED SPEAKER / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 108 JOSHUA L. WORT1,2, TO BIND OR NOT TO BIND – THE SLINGS AND ARROWS KATRIN ACKERMANN1,2, OF BIOMOLECULAR BINDING STUDIES BY PULSE MARIA ORANGES3, LAURA REMMEL1,2, DIPOLAR EPR SPECTROSCOPY. ANGELIKI GIANNOULIS3, ALAN J. STEWART2,4, Pulse dipolar electron paramagnetic resonance spectroscopy (PDS) has become an increasingly popular tool to BELA E. BODE1,2 investigate topologies and conformational flexibilities of biomacromolecules. In addition to mere distance information quantitative PDS gives access to the number of interacting spins. Early model studies demonstrate that 1School of Chemistry, University of St Andrews, St Andrews, Scotland, UK discrete multimeric states [1], dimerisation equilibria [2], metal ion binding site occupation [3], and cooperativities 2Biomedical Sciences Research of binding [4] can be determined. We have recently exploited this to demonstrate the surprisingly efficient [5, 6] and Complex and Centre of Magnetic robust [7] coordinative copper(II) based spin labelling of double histidine motifs in proteins offering highly precise Resonance, University of St Andrews, St Andrews, Scotland, UK distance measurements at nanomolar cooncentrations [8]. 3Weizmann Institute of Science, Rehovot, Israel Two examples highlighting the level of detail that can be extracted from quantitative PDS will be presented. A sin-4School of Medicine, University of St gle-stranded DNA binding protein [9] forming dimers and dimers of dimers, and a templated dimerisation of a Andrews, St Andrews, Scotland, UK model protein both allow extracting multiple parameters characterising the equilibria from PDS data. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS References: [1] B. E. Bode, D. Margraf, J. Plackmeyer, G. Durner, T. F. Prisner, O. Schiemann . , J. Am. Chem. Soc. 2007, 129, 6736-6745. [2] K. Ackermann, A. PRIZE LECTURES Giannoulis, D. B. Cordes, A. M. Z. Slawin, B. E. Bode, Chem Commun. 2015, 51, 5257-5260. [3] A. Giannoulis, M. Oranges, B. E. Bode, ChemPhysChem. 2017, PLENARY LECTURES 18, 2318-2321. [4] A. Giannoulis, K. Ackermann, P. Spindler, C. Higgins, D. B. Cordes, A. M. Z. Slawin, T. F. Prisner, B. E. Bode, Phys. Chem. Chem. Phys. 2018, TUTORIAL LECTURE 20, 11196-11205. [5] J. L. Wort, K. Ackermann, A. Giannoulis, A. J. Stewart, D. G. Norman, B. E. Bode, Angew. Chem. Int. Ed. 2019, 58, 11681-11685. [6] J. L. Wort, K. Ackermann, D. G. Norman, B. E. Bode, Phys. Chem. Chem. Phys. 2021, 23, 3810-3819. [7] J. J. L. Wort, S. Arya, K. Ackermann, A. J. Stewart, B. E. Bode, J. Phys. INVITED AND PROMOTED LECTURES Chem. Lett. 2021, 12, 2815-2819. [8] K. Ackermann, J. L. Wort, B. E. Bode, J. Phys. Chem. B 2021, 125, 5358-5364. [9] M. J. Morten, J. R. Peregrina, M. Figueira- POSTERS Gonzalez, K. Ackermann, B. E. Bode, M. F. White, J. C. Penedo, Nucleic Acids Res. 2015, 43, 10907-10924. ADVERTISERS Acknowledgements: We are indebted to the Leverhulme Trust (RPG-2018-397) and the Wellcome Trust (204821/Z/16/Z) for funding. We gratefully AUTHOR INDEX acknowledge equipment funding through the Wellcome Trust (099149/Z/12/Z) and BBSRC (BB/R013780/1). PT040 PARALLEL SESSIONS / PROMOTED TALK / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 109 THOMAS SCHMIDT, ACCESSING CALMODULIN BINDING NONEQUILIBRIUM JAEYKUN JEON, KINETICS BY EPR DIPOLAR SPECTROSCOPY ROBERT TYCKO, G MARIUS CLORE As the cell’s prototypical Ca2+ sensor, calmodulin (CaM) is responsible for amplifying and adapting this universal sigNational Institutes of Health, nal into specific downstream signals via interactions with numerous target proteins in a calcium-dependent man- Laboratory of Chemical Physics, ner. This biological adaptability correlates with the significant structural plasticity that has been observed in CaM. Bethesda, USA Here, the interaction of CaM and its 26-residue binding partner, M13, was probed on the millisecond time scale. The interaction mechanism is well established and, hence, serves as a model to evaluate the nonequilibrium kinetics by EPR dipolar spectroscopy. Our method uses a combination of selective protonation/deuteration [1], rapid mix and subsequent rapid freeze-quench [2] along the CaM and M13 reaction coordinates. Further, the acquisition of T -ed-m ited DEER [3] and subsequent processing by 2D SF-SVD enable detailed description of conformational components. The interaction between CaM and M13 gives rise to two intermediates originating from the initial interaction event (2 ms) and the coordination of the N- and C-terminal domains (8 ms) in addition to its halo- and fully bound state. The presented technique can be expanded to a wide range of molecular systems to answer questions of protein folding and domain coordination. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] R. Ward, A. Bowman, E. Sozudogro, H. El-Mkami, T. Owen-Hughes, D. Norman JMR. 2010, 207, 164-167. [2] T. Schmidt, J. Jeon, Y. Okuno, S.C. AUTHOR INDEX Chiliveri G.M. Clore Chem. Phys. Chem. 2020, 120, 356-365. [3] T. Schmidt, G.M. Clore, Chem. Com. 2020, 56, 10890-10893. PT041 PARALLEL SESSIONS / PROMOTED TALK / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 110 ELENA G. BAGRYANSKAYA1, IVAN O. DEER/PELDOR STUDY OF SUPRAMOLECULAR TIMOFEEV2,3, KONSTANTIN N. BULYGIN 4, ASSEMBLIES OF HUMAN RIBOSOME AND RNAS ALEXEY A. MALYGIN 3,4, DMITRI M. GRAIFER 3,4, MARIA I. MESCHANINOVA4, Pulse dipolar EPR was used to investigated structural rearrangements in mRNA upon its binding to human 80S ribosomes ALYA G. VENYAMINOVA4, OLESYA A. [1,2,3]. The model mRNA (MR), 11-mer RNA containing two nitroxide spin labels at the 5 - and 3 -terminal nucleotides and KRUMKACHEVA2,3, MATVEY V. FEDIN2,3, prone to form a stable homodimer (MR)2, was used. Intramolecular spin-spin distances were measured by DEER/PELDOR LUDMILA YU. FROLOVA5, GALINA G. spectroscopy in model complexes mimicking different states of the 80S ribosome during elongation and termination KARPOVA4 of translation. The formation of two different types of ribosomal complexes with MR was observed [1]. First, there were 1N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry SB stable complexes where MR was fixed in the ribosomal mRNA-binding channel by the codon-anticodon interaction(s) RAS, Novosibirsk, Russia 2 with cognate tRNA(s). Second, we for the first time detected complexes assembled without tRNA due to the binding of International Tomography Center SB RAS, Novosibirsk, Russia 3Novosibirsk State University, Novosibirsk, Russia MR most likely to an exposed peptide of ribosomal protein uS3 away from the mRNA channel. Our findings showed that a 4Institute of Chemical Biology and Fundamental Medicine SB part of mRNA bound in the ribosome channel, which is not involved in codon-anticodon interactions, has more degrees RAS, Novosibirsk, Russia 5 of freedom than that interacting with tRNAs. The features of labile complexes of human 40S ribosomal subunits with Engelhardt Institute of Molecular Biology RAS, Moscow, Russia RNAs, whose formation is manifested in the cross-linking of aldehyde derivatives of RNAs to the ribosomal protein uS3 through its peptide 55–64 located outside the mRNA channel was studied in details [2]. The measurements revealed that in all studied complexes, mRNA exists in two alternative conformations, whose ratios are different in post-translocation, pre-translocation and termination complexes. We found that the presence of aminoacyl-tRNA at the ribosomal A site decreases the relative share of the more extended mRNA conformation, whereas the binding of eRF1 (alone or in a complex with eRF3) results in the opposite effect. In the termination complexes, the ratios of mRNA conformations are practically INTRODUCTION the same, indicating that a part of mRNA bound in the ribosome channel does not undergo significant structural alter- SPONSORS ations in the course of completion of the translation. Our results contribute to the understanding of mRNA molecular PROGRAM dynamics in the mammalian ribosome channel during translation [3]. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] A.A. Malygin, O.A. Krumkacheva, D.M. Graifer, et al. E.G. Bagryanskaya, NAR 2019, 47, 11850-11860, [2] A.A. Malygin, et al. NAR, 2018, 46, POSTERS 897-904. [3] K.N. Bulygin, I. O.Timofeev, A.A. Malygin, D.M. Graifer, M.I. Meschaninova, A.G. Venyaminova, O.A. Krumkacheva, M.V. Fedin, L.Yu. Frolova, ADVERTISERS G.G. Karpova, E.G. Bagryanskaya, Computational and Structural Biotechnology Journal, 2021, in press. AUTHOR INDEX Acknowledgements: This work was supported by Russian Ministry of Science and Higher Education within Grant 14.W03.31.0034 (megagrant). PT042 PARALLEL SESSIONS / PROMOTED TALK / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 111 YVO POKERN1, STATISTICAL ANALYSIS OF ENDOR AT 263 GHZ BENJAMIN ELTZNER2, REVEALS A CONFORMATIONAL DISTRIBUTION IN A STEPHAN HUCKEMANN2, CLEMENS BEEKEN2, PROTEIN TYROSYL RADICAL IGOR TKACH3, MARINA BENNATI3, Electron nuclear double resonance (ENDOR) spectroscopy probes the hyperfine (hf) interaction of unpaired elec- MARKUS HILLER3 trons and magnetic nuclei in their environment, yielding structural information at atomic resolution. Technical 1 advancements have enabled ENDOR spectroscopy at increasingly larger magnetic field strengths, recently extend- Department of Statistical Science, University College London, London, ing to 9.4 T (263 GHz) [1]. The spectra recorded in this study on the Y • radical in E. Coli ribonucleotide reductase 122 United Kingdom suggested the presence of a hf interaction, which had previously been overlooked. The unusually broad appearance 2Department of Mathematical of this signal prompted us to develop a new approach for processing ENDOR data that allows for signals to be distin-Stochastics, Georg-August-University Göttingen, Göttingen, Germany guished from baseline distortions [2]. The method is based on a mathematical modelling of the measurement sig- 3Max Planck Institute for Biophysical nal considering variations between individual scan repetitions and a processing scheme to obtain the most-likely Chemistry, Göttingen, Germany ENDOR signals. Furthermore, an estimation of confidence intervals for the spectrum becomes feasible. Application of this new method in combination with additional statistical tests confirms the presence of broad resonances in the Y • ENDOR spectra in a quantitative way based on p values. We anticipate that our approach can be generally 122 applied to ENDOR spectroscopy and will ultimately allow the quantification of uncertainties of spin density distributions and structural information that can be extracted from ENDOR data [2,3]. Isotopic labelling experiments were employed, which unambigously linked the newly recognized hf coupling to INTRODUCTION the β-methylene group of the radical. Finally, the unusually broad shape of these resonances could be explained SPONSORS by spectral simulations in conjunction with density functional theory by assuming a normal distribution of ring PROGRAM dihedral angles, which is known to strongly influence the hf coupling of β-methylene protons [4]. This study pro- PROGRAM — BY DAY vides a new approach for analyzing structural inhomogeneity in tyrosyl radials, which are essential for a variety on TOPICS biological systems. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] I. Tkach, I. Bejenke, F. Hecker, A. Kehl, M. Kasanmascheff, I. Gromov, I. Prisecaru, P. Höfer, M. Hiller, M. Bennati, J. Magn. Res. 2019, 303, 17-27. ADVERTISERS [2] Y. Pokern, B. Eltzner, S. Huckemann, C. Beeken, J. Stubbe, I. Tkach, M. Bennati, M. Hiller, PNAS, accepted. [3] S. Pribitzer, D. Mannikko, S. Stoll, Phys Chem AUTHOR INDEX Chem Phys 2021, 23, 8326-8335. [4] C. Heller, H. M. McConnell, J. Chem. Phys. 1960, 32, 1535-1539. PT043 PARALLEL SESSIONS / PROMOTED TALK / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 112 MAXIMILIAN GAUGER,1 INVESTIGATION OF THE CONFORMATIONAL DYNAMICS OF DSRNA LUKAS S. STELZL,2 USING ORIENTATION-SELECTIVE PELDOR AND MD SIMULATIONS ANNA-LENA J. SEGLER,3 NICOLE ERLENBACH,1 AS COMPLEMENTARY METHODS GERHARD HUMMER4, SNORRI TH. SIGURDSSON3 AND DNA and RNA form the basis for storage and transmission of genet- spin labels and with that insights into the dsRNA helix geometry THOMAS F. PRISNER1 ic information in living organisms. DNA carries the genetic infor- and dynamics can be gained. This is, however, not a straightforward 1 mation which is transcribed to RNA. Single- and double-stranded task and may yield ambiguous results. Here, we used conformers Institute of Physical and Theoretical Chemistry and Center of Biomolecular Magnetic Resonance, RNA (ssRNA and dsRNA) is needed for protein biosynthesis. Apart predicted by molecular dynamics (MD) simulations using different Goethe University Frankfurt, Frankfurt am Main, from their function as carriers of genetic information, nucleic acids force fields to calculate orientation-selective PELDOR time traces. Germany 2 also play a crucial role in the regulation of cellular processes. In all Institute of Physics, Johannes Gutenberg Experimental orientation-selective PELDOR data were acquired at University, Mainz, Germany these tasks, the conformational dynamics of the nucleic acids can 3 X- (9.5 GHz, 0.3 T) and G-band (180 GHz, 6.4 T) where differing de- Science Institute, University of Iceland, be ascribed as the root cause of their function. Reykjavik, Iceland grees of orientation-selection are achievable. Using simulated data 4Department of Theoretical Biophysics, Max The aim of this project was the investigation of the conformational from MD predictions using the DESRES [2] force field we were able Planck Institute of Biophysics, Frankfurt am Main, Germany dynamics of dsRNA. The system we chose for these studies was a to achieve very good agreement between experimental and simu-set of 20-mer dsRNA molecules in A-helix form. Each dsRNA mole- lated data. This way we validated the accuracy of the DESRES force cule was doubly labelled with the rigid Çm spin label [1] in varying field for the dsRNA under investigation and can proceed with the positions. Pulsed ELectron Double Resonance (PELDOR) spectros-analysis of the simulated data. It will be interesting to analyze the copy time traces are modulated by the electron-electron dipolar helix geometry and its conformational dynamics and compare our INTRODUCTION coupling. Due to the rigidity of the Çm spin label it is possible to findings to previously reported findings for dsDNA [3] and dsRNA SPONSORS obtain sets of orientation-dependent data using multi-frequency/ [4]. PROGRAM multi-field orientation-selective PELDOR experiments. From such PROGRAM — BY DAY data insights about the mutual orientation and distances of the TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] C. Hörbartner, G. Sicoli, F. Wachowius, D.B. Gophane, S.Th. Sigurdsson, J. Org. Chem 2012, 77, 7749-7754. [2] D. Tan, S. Piana, R.M. Dirks, D.E. Shaw, Proc. Natl. Acad. Sci. USA POSTERS 2018, 115, E1346-E1355. [3] L.S. Stelzl, N. Erlenbach, M. Heinz, T.F. Prisner, G. Hummer, J. Am. Chem. Soc. 2017, 139, 11674-11677. [4] K. Liebl, T. Drsata, F. Lankas, J. Lipfert, M. Zacharias, Nucleic ADVERTISERS Acids Res. 2015, 43, 10145-10156. AUTHOR INDEX Acknowledgements: We acknowledge financial support from the German Research Foundation (CRC 902: Molecular Principles of RNA based regulation). IN033 PARALLEL SESSIONS / INVITED SPEAKER / DRUG DESIGN AND COMBAT AGAINST COVID-19 113 GYULA PÁLFY1, UNDERSTANDING ONCOGENIC KRAS CYCLE THROUGH ORSOLYA TŐKE2, MULTIPLE TIME-SCALE DYNAMICS ISTVÁN VIDA1, DÓRA K. MENYHÁRD1, KRas (Kirsten RAt Sarcoma virus) signaling protein is inactive when GDP occupies the binding site. Followed by HANNA ÁKONTZT-KISS1, the GDP to GTP exchange, a process catalyzed by GEF (Guanine nucleotide Exchange Factors) proteins, the spatial GYULA BATTA3, structure of the GTP binding protein changes and thus, becomes active in signal transduction. KRas GTP hydrolysis ANDRÁS PERCZEL1 is accelerated by GAPs (GTPase Activating Proteins). Once GTP hydrolysis and there will be GDP again in the binding 1MTA-ELTE Protein Modeling Research pocket, 3D-fold changes again and the protein will return to its off-state mode. kRas protein consists of two parts: Group & Laboratory of Structural the effector and the allosteric regions. As a result of our comprehensive Lipari-Szabo model-free and reduced spec-Chemistry and Biology, Eötvös Loránd University, Budapest, Hungary tral density mapping plus CPMG analysis on the wide type and 3 mutants, G12C, G12D and G12V, we found that both 2Laboratory for NMR Spectroscopy, kRas.GTP.Mg2+, kRas.GDP.Mg2+ has a major- and a minor-form. However, the latter on, crucial in understanding the Research Center for Natural Sciences details of GDP to GTP exchange, appears to be an on-pathway intermediate state of the transition. To characterize a (RCNS), Budapest, Hungary 3Department of Organic Chemistry, minor form is a major challenge, but we found that the 3D-structure of the Mg2+-free state of kRas shows high sim-University of Debrecen, Debrecen, ilarity to the minor-form. Hungary. 1) We found that the relative distances of the P-loop, switch-I, and switch-II increase in the Mg2+-free state, respectively, as they do in the X-ray determined (GDP.Mg2+)free KRas.GEF complex. 2) We observed that in the case of the wt, G12C and G12D mutants, the distance between selected residue pairs of the β2-β3 antiparallel strands increases in the Mg2+-free-state. β-strands do separate and move away from each other. If we superimpose the wt, G12V and wt(GDP.Mg2+)free kRas.GEF 3D-structures, then will be clear that the bulge described above has functional signifi- INTRODUCTION cance. This bulge will host GEF protein when it forms a complex with kRas to facilitate the release of bound GDP. 3) SPONSORS We found that the strength of the H-bonds formed between HO2’- and HO3’ of the furanose ring of GDP and adja- PROGRAM cent C=Os (V29 and D30) changes with the Mg2+ ion release. We may ask how relevant all these data are for drug dis- PROGRAM — BY DAY covery? It would be good to design and then synthesize small molecules that are able to bind the herein described TOPICS and characterized minor-form of kRas mutants and thus remove them selectively. The selectively elimination of PRIZE LECTURES harmful KRas mutants could stop abnormal signaling and thus, have the potential to cure tumor cells. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] Gy. Pálfy, D. K. Menyhárd, A. Perczel Cancer and Metastasis Reviews 2020. [2] D. K. Menyhárd, Gy. Pálfy, Z. Orgován, I. Vida, Gy. M. Keserű, ADVERTISERS A.Perczel, Chemical Science 2020. [3] Gy. Pálfy, I. Vida, A. Perczel, Biomolecular NMR Assignments 2020, 14, 1-7. [4] M. Rachman, A. Scarpino, D. Bajusz, Gy. AUTHOR INDEX Pálfy, I. Vida, A. Perczel, X. Barril, Gy. M. Keserű, ChemMedChem 2019, 14, 1-12. IN034 PARALLEL SESSIONS / INVITED SPEAKER / DRUG DESIGN AND COMBAT AGAINST COVID-19 114 MAVROMOUSTAKOS NMR AND RATIONAL DRUG DESIGN THOMAS Nuclear Magnetic Resonace (NMR) spectroscopy is considered the most powerful analytical method as it finds National Kapodistrian University of Athens, Chemistry plethora of applications in various fields including chemistry, biology, pharmacy and medicine. One of these ap-Department, Organic Chemistry plications that combines the use of the above mentioned fields is the “Rational Drug Design”. Drugs now are now Laboratory, Athens, Greece designed rationally using in silico techniques combined with analytical methods such as liquid and solid state NMR spectroscopy based on the existing molecular basis of information. In our laboratory we are using this combined methodology for studying different aspects of rational drug design. In particular we: (i) explore the conformational space of commericially available drug molecules in order to discover novel structures using a combination of NMR spectroscopy and computational chemistry and especially Molecular Dy- namics calculations; (ii) study the interactions of drugs with their vehicles such as cyclodextrins and calixarenes in an attempt to relate their biological properties and strength of their physical chemical interactions; Again, Molecular Docking, Molecular Dynamics and 3D Quantitative Structure Activity Relationships (3D QSAR) provide complementary information. (iii) investigate the interactions of drug molecules in lipid bilayers in order to reveal the membrane role in their biological action; (iv) examine the drug stability. INTRODUCTION In the talk examples will be provided from our recent work which will illustrate that: SPONSORS (i) NMR spectroscopy can provide a valuable information in the rational drug design; PROGRAM (ii) It can be combined fruitfully with computational analysis to enhance the knowledge on drug action and un- PROGRAM — BY DAY derstand aspects related to molecular basis of their action. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] S. Kiriakidi, C. Chatzigiannis, C. Papaemmanouil, A.G. Tzakos, T. Mavromoustakos. Biochimica et Biophysica Acta 2020, 1862, 183142. [2] K. Manta, P. Papakyriakopoulou, M. Chountoulesi, D.A. Diamantis, D. Spaneas, V. Vakali, N. Naziris, M.V. Chatziathanasiadou, I. Andreadelis, K. Moschovou, POSTERS I. Athanasiadou, P. Dallas, D.M. Rekkas, C. Demetzos, G. Colombo, S. Banella, U. Javornik, J. Plavec, T. Mavromoustakos, A.G. Tzakos, G. Valsami, Molecular ADVERTISERS Pharmaceutics 2020, 17(11), 4241-4255. [3] S. Kiriakidi, C. Chatzigiannis, C. Papaemmanuouil, A.G. Tzakos, Z. Cournia, T. Mavromoustakos, Computational and AUTHOR INDEX Structural Biotechnology Journal 2021 19, 110–120. [4] A. Chroni, T. Mavromoustakos, S. Pispas. Polymers 2021, 13(7), 1164. IN035 PARALLEL SESSIONS / INVITED SPEAKER / DRUG DESIGN AND COMBAT AGAINST COVID-19 115 ANTONIO TARGETING G-QUADRUPLEX AND I-MOTIF DNA STRUCTURES: RANDAZZO A NEW OPPORTUNITY IN ANTI-CANCER THERAPY University of Naples Federico II, Department of Parmacy, Naples, The most widely recognized genomic DNA structure is the classical also been found to be involved in telomere biology and in the reg-Italy DNA double helix. However, DNA is structurally very dynamic and ulation of gene-transcription [1]. Interestingly, the formation of able to adopt several alternative secondary structures, such as cru-iM structures is cell-cycle dependent, peaking at late G1 phase [5], ciforms, G-quadruplexes, triplexes, and i-motifs [1]. G-quadruplex whereas G4 formation is maximal during S phase [6]. This suggests structures (G4s) are among the most extensively studied DNA sec-that iMs and G4s might play different roles in regulating gene ex- ondary structures [2]. They can be unimolecular or intermolecular pression and transcription and potentially their presence could be and can adopt a wide diversity of topologies depending on the com-mutually exclusive. Big efforts have been made by the international binations of strand orientation and length and composition of the scientific community to find and develop small molecules that can loops. G4 structures are formed by guanine-rich sequences that are selectively recognize G4 and iM structures, aiming at disentangling located mainly in telomeres, gene promoters and in the first intron the individual biological processes regulated by these structures in of genes [3], where they are involved in the regulation of key biolog-the context of cancer cells. Many molecules display considerable ical processes, like, for example, telomere protection, regulation of selectivity for G4s over single-stranded and double-stranded DNA, transcription, and translation. and some of them are also capable to discriminate among G4 fold- ing topologies, generally distinguishing parallel from anti-paral- The G4-forming complementary strand is a single stranded C-rich lel quadruplexes. Very interestingly, many G4 ligands were found sequence, potentially bound by single-stranded binding proteins [4]. to exhibit intriguing anti-cancer activities. Differently from the Alternatively, the C-rich strand might fold in another four-stranded INTRODUCTION well-documented examples of G4 ligands, the discovery of specific structure called i-motif (iM). This is composed of two head-to-tail, SPONSORS iM ligands lags far behind. In this communication examples of G4 intercalated, parallel-stranded duplexes held together by hemi-pro- PROGRAM and iM ligands will be shown. tonated cytosine-cytosine+ (C·C+) base pairs. iM structures have PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] H.A. Assi, M. Garavís, C. González, M.J. Damha, Nucleic Acids Res 2018, 46(16), 8038–8056. [2] J. Spiegel, S. Adhikari, S. Balasubramanian, Trends Chem 2019:1–14. [3] G. INVITED AND PROMOTED LECTURES Marsico, V.S. Chambers, A.B. Sahakyan, P. McCauley, J.M. Boutell, M. Di Antonio, et al. Nucleic Acids Res 2019, 47(8), 3862–3874. [4] S.L. Brown, S. Kendrick, Pharmaceuticals 2021, 14(2), 96. POSTERS [5] M. Zeraati, D.B. Langley, P. Schofield, A.L. Moye, R. Rouet, W.E. Hughes, et al. Nat Chem 2018;10(6):631–637. [6] G. Biffi, D. Tannahill, J. McCafferty, S. Balasubramanian Nat Chem 2013, ADVERTISERS 5(3), 182–186. AUTHOR INDEX Acknowledgements: Italian Association for Cancer Research (AIRC) is aknowledge for finantial support. PT044 PARALLEL SESSIONS / PROMOTED TALK / DRUG DESIGN AND COMBAT AGAINST COVID-19 116 SEBASTIAN HILLER THE OUTER MEMBRANE INSERTASE AS Biozentrum, University of Basel, AN ANTIBIOTIC TARGET Basel, Switzerland Proteins in the outer bacterial membrane are attractive antibiotic targets, because their location in the cellular pe-riphery prevents many bacterial counteracting strategies. The outer membrane insertase BAM, one of two generally essential outer membrane proteins, is such a target. BAM receives its membrane protein substrates from a chain of periplasmic chaperones and then folds and inserts them into the membrane. BAM lacks a classical catalytic center and for long time it remained questionable whether BAM can be inhibited at all. I will describe an integrated structural biology approach to resolve the functional mechanism of the BAM insertase in outer membrane protein biogenesis and to demonstrate that two recently discovered peptidic antibiotics, the natural compound darobactin and the engineered bicyclic OMPTA, inhibit BAM by direct interaction [1,2]. Solution NMR studies based on extensive sequence-specific resonance assignments of the BAM transmembrane barrel [3], combined with cryo-electron microscopy, X-ray crystallography, biophysics and molecular dynamics simulation show that darobactin inhibits the BAM complex by mimicking the cognate substrate recognition sequence [4]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] A. Luther, M. Urfer, et al., Nature 2019, 576, 452-458. [2] Y. Imai, K.J. Meyer, et al. Nature. 2019, 576, 459-464. [3] J.B. Hartmann et al., J. Am. AUTHOR INDEX Chem. Soc. 2018, 140, 11252-11260. [4] H. Kaur, R.P. Jakob et al., Nature. 2021, available online. IN036 PARALLEL SESSIONS / INVITED SPEAKER / DRUG DESIGN AND COMBAT AGAINST COVID-19 117 HARALD SCHWALBE INVESTIGATING THE PROTEOME AND on behalf of the GENOME OF SARS-COV-2 Covid19-nmr Consortium SARS-CoV-2 contains a positive single-stranded RNA genome of approximately 30,000 nucleotides coding for ~ Center for Biomolecular 27 different proteins. Within this genome, 15 RNA elements were identified as conserved between SARS-CoV and Magnetic Resonance (BMRZ), SARS-CoV-2. By nuclear magnetic resonance (NMR) spectroscopy, we determined that these elements fold inde- Goethe-University Frankfurt, Frankfurt, Germany; pendently and provided 1H, 15N chemical shift assignments for all these elements [1]. We optimized expression con-schwalbe@nmr.uni-frankfurt.de ditions for 24 proteins, conducted NMR experiments to establish folding conditions [2] and assigned more than 10 of these proteins within covid19-nmr [3]. We performed NMR-based screenings with a poised fragment library of 768 compounds for binding to these viral RNAs [4] and proteins [5] employing three different 1H-based 1D NMR binding assays. Initial hits have been analyzed by cheminformatics approaches and follow-up strategies are being discussed for selected RNA and protein examples. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1]Wacker, Weigand et al. Nucl. Acids Res. 2020, 48, 12415-35. [2] Altincekic, Korn, Qureshi, Dujardin, Ninot-Pedrosa et al. Front Mol Biosci. 2021, ADVERTISERS 8, 653148. [3] For updates on chemical shift assignments, see covid19-nmr.de. [4] Sreeramulu, Richter et al. Angew. Chem. 2021, in revision. [5] Manuscript AUTHOR INDEX in preparation. IN037 PARALLEL SESSIONS / INVITED SPEAKER / DRUG DESIGN AND COMBAT AGAINST COVID-19 118 ANA ARDÁ, STRUCTURAL CHARACTERIZATION OF N-LINKED JUNE-EREÑO-ORBEA, GLYCANS IN THE RECEPTOR BINDING DOMAIN JESÚS JIMÉNEZ-BARBERO Chemical Glycobiology lab, CIC bioGUNE, OF THE SARS-COV-2 SPIKE PROTEIN AND THEIR Basque Research & Technology Alliance (BRTA) INTERACTIONS WITH HUMAN LECTINS Ikerbasque, Basque Foundation for Science, Bizkaia, Spain The glycan structures of the receptor binding domain of the SARS-CoV2 spike glycoprotein expressed in human HEK293F cells have been studied by using NMR. The different possible interacting epitopes have been deeply an- alysed and characterized, providing evidence of the presence of glycan structures not found in previous MS-based analyses. The interaction of the RBD 13C-labelled glycans with different human lectins, which are expressed in different organs and tissues that may be affected during the infection process, has also been evaluated by NMR. In particular, 15N-labelled galectins (galectins-3, -7 and -8 N-terminal), Siglecs (Siglec-8, Siglec-10), and C-type lectins (DC-SIGN, MGL) have been employed. Complementary experiments from the glycoprotein perspective or from the lectin’s point of view have permitted to disentangle the specific interacting epitopes in each case. Based on these findings, 3D models of the interacting complexes have been proposed. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN038 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULAR DYNAMICS 119 PETRA ROVÓ1,2 R RELAXATION DISPERSION UNDER FAST 1R 1Ludwig Maximilian University, MAGIC-ANGLE SPINNING Munich, Germany 2Institute for Science and Technology Austria, Recent advances in instrumentations, isotope-labeling schemes, sequence of squid ring teeth proteins [2]. Hydration-inducedline Klosterneuburg, Austria and the continued method developments for fast magic-an- narrowing in solid-state NMR spectra is a known phenomenon for gle-spinning (MAS) applications provide unprecedented insights soft biopolymer materials, such as elastin, collagen, silk. Enhanced into biologically relevant structural dynamics of large protein commotion on the nanosecond timescale can beprobed by 13C longitudi- plexes or insoluble biomacromolecules. The importance of micro- nal relaxation, while changes in microsecond timescale dynamics second time-scale conformational exchange in protein-protein isbestassessed using 13C rotating-frame relaxation dispersion tech-interaction, allostery, or signal transduction is widely recognized niques. In particular, I will demonstrate how 13C off-resonance Near and extensively studied with solution-state NMR techniques; with Rotary-Resonance Relaxation Dispersion (NERRD) method can be solid-state NMR, such explorations are just about to unravel. In this used to identify the origin and quantify the amplitude of the fast talk, I briefly summarize the theoretical aspects and the practical microsecond time-scale motion. The application of fast magic-an-considerations of solid-state magic-angle-spinning rotating-frame gle spinning (55.55 kHz) and increasing spin-lock field strengths relaxation dispersion methods that are particularly sensitive to fast approaching the half-and full-rotary-resonance conditions enables microsecond structural transitions. Unlike in solution state, in sol-the separatoin of the relaxation dispersion regime that reports id-state MAS measurementsrotating-frame (R ) relaxation disper- on incoherent dynamics as opposed to coherent dephasing of the 1r sion can be monitored at two distinct effective radio-frequency field transverse magnetization due to the presence of dense proton net- regimes (close to zero frequency and close to the rotary-resonance work. These relaxation measurements revealed that the motion of INTRODUCTION conditions) delivering complementary information about fast con- proline side-chain carbons are the most affected by hydration giv- SPONSORS formational exchange processes [1]. ing rise to sharp signals, and hence to long coherence life times, in PROGRAM both dipolar-coupling-based and scalar-coupling-based 2D 1H–13C As an example, I will demonstrate the applicability of the method to PROGRAM — BY DAY experiments. study the structural and dynamic transitions of a de novodesigned TOPICS soft polymer materialwhose amino acid sequence is based on the PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] P. Rovó, Solid State Nucl. Magn. Reson. 2020, 108, 101665. [2] R. C. A. Dubini, H. Jung, M. C. Demirel, P. Rovó, J. Phys. Chem. B. 2021, 125, 2134-2145. IN039 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULAR DYNAMICS 120 LORENZO BARONTI1, BASE-PAIR SWITCHING MODULATES MICRORNA ILEANA GUZZETTI1, ACTIVITY PARISA EBRAHIMI2, SARAH FRIEBE SANDOZ1, Many functions of RNA depend on rearrangements in secondary structure that are triggered by external factors, EMILIE STEINER1, such as protein or small molecule binding. These transitions can feature on one hand localized structural changes JUDITH SCHLAGNITWEIT1, in base-pairs or can be presented by a change in chemical identity of e.g. a nucleobase tautomer. We use and de-BASTIAN FROMM3, velop R -relaxation-dispersion NMR method [1] for characterizing transient structures of RNA that exist in low 1r LUIS SILVA1, abundance (populations <10%) and that are sampled on timescales spanning three orders of magnitude (µs to s). CAROLINA FONTANA1, ALAN A. CHEN2, The characterization of microRNA 34a targeting the p53 modulator mRNA of Sirt1. This microRNA – mRNA com- KATJA PETZOLD plex changes conformation to activate the RISC complex [2] (Nature 2020). I will furthermore give an outlook on 1 recent efforts to measure in-cell NMR of nucleic acids in functional complexes [3]. Dept. of Med. Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden 2Department of Chemistry and RNA Institute, University at Albany, State University of New York, Albany, NY, USA 3Science for Life Laboratory, Dept. of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] Steiner, E., Schlagnitweit, J., Lundström, P. & Petzold, K. Angewandte. 2016, 55 (51), 15869–15872 / J. Schlagnitweit, E. Steiner, H. Karlsson & TUTORIAL LECTURE K. Petzold, Chemistry – A European Journal. 2018 24(23):6067-6070; [2] L. Baronti, I. Guzzetti, P. Ebrahimi, S. Friebe Sandoz, E. Steiner, J. Schlagnitweit, B. Fromm, L. Silva, C. Fontana, A. A. Chen & K. Petzold Nature 2020, 583, 139–144; [3] J. Schlagnitweit, S. Friebe Sandoz, A. Jaworski, I. Guzzetti, F. Aussenac, R. INVITED AND PROMOTED LECTURES J. Carbajo, E. Chiarparin, A. J. Pell & K. Petzold, ChemBioChem. 2019, 20(19): 2474-2478; www.petzoldlab.com POSTERS Acknowledgements: N. Hopkins, E.R. Andersson, Petzold Lab members, I. MacRae, Lund University Protein Production Platform, Protein Science Facility. ADVERTISERS K.P. acknowledges funding from the Swedish Research Council, the Swedish Foundation for Strategic Research, Harald och Greta Jeansson Stiftelse, Carl AUTHOR INDEX Tryggers stiftelse Eva och Oscar Ahréns Stiftelse, Åke Wiberg Stiftelse, Cancerfonden, Ragnar Söderberg Stiftelse (M91/14). IN040 PARALLEL SESSIONS / INVITED SPEAKER / BIOMOLECULAR DYNAMICS 121 FALK HOFFMANN1, NARROWING THE GAP BETWEEN NMR RELAXATION AHMED A.A.I. ALI1, AND MD SIMULATION STUDIES OF PROTEIN MENGJUN XUE2, LARS V. SCHÄFER1, DYNAMICS FRANS A.A. MULDER2 1 While NMR nuclear spin relaxation is arguably the most comprehensive experimental approach to measure protein Theoretical Chemistry, Ruhr University Bochum, Bochum, dynamics at atomic resolution, it is only highly approximate due to the very sparse sampling of the spectral density Germany function (SDF) it provides. MD simulation, on the other hand, describe molecular flexibility in amazing detail, but 2 Interdisciplinary Nanoscience are constrained by limitations on the accuracy of the molecular mechanics force field models and conformational Center (iNANO) and Department of Chemistry, Aarhus University, sampling. The two methods are therefore highly complementary for the study of picosecond to nanosecond time Aarhus, Denmark scale dynamics in proteins, and can act in a highly synergistic manner [1]. We find that the popular Lipari-Szabo ‘model free’ approximation of the internal time correlation functions is highly inadequate [2]. Instead, an objective comparison is feasible if we use the full MD trajectories to compute the SDF, including anisotropic molecular tumbling [2,3]. Examples of 13C and 2H NMR relaxation data will be provided and discussed [2-4]. The decreasing gap between prediction and experiment points to quantitative avenues for probing configurational entropy [5,6]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Hoffmann F., Mulder F. A. A., Schäfer L. V. J Phys Chem B. 2018, 122(19):5038-5048. [2] Hoffmann F., Xue M., Schäfer L. V., Mulder F. A. A., Phys INVITED AND PROMOTED LECTURES Chem Chem Phys. 2018, 20(38):24577-24590. [3] Hoffmann F., Mulder F. A. A., Schäfer L. V. J Chem Phys. 2020, 152(8):084102. [4] Ali, A. A. I., Hoffmann F., POSTERS Schäfer L. V., Mulder F. A. A., 2021 in preparation. [5] Chakravorty A., Higham J., Henchman R. H. J Chem Inf Model. 2020, 60(11):5540-5551. [6] Hoffmann F., ADVERTISERS Mulder F. A. A., Schäfer L. V, 2021 in preparation. AUTHOR INDEX Acknowledgements: We thank the RESOLV cluster of excellence at RU Bochum and the Center for Ultra High Field NMR Spectroscopy at Aarhus University. PT045 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULAR DYNAMICS 122 ALBERT A. SMITH, UNVEILING THE DYNAMIC LANDSCAPE WITH NMR RELAXATION ALEXANDER VOGEL, AND MOLECULAR DYNAMICS SIMULATION OSKAR ENGBERG, PETER HILDEBRAND, Biomolecular function is the result of a complex hierarchy of mo- motion as a function of molecular position and correlation time. DANIEL HUSTER lecular motions. NMR relaxation is one of the premier experimen- While we have applied our method to POPC membranes, our ap- Leipzig University, Institute for tal methods for characterization of dynamics, having an array of proach is fully general and may be applied to any system for which Medical Physics and Biophysics, timescale-selective and bond-specific experiments available for the experimental and simulated data is available. Leipzig, Germany characterization of reorientational motion. Still, these experiments capture the aggregate influence of multiple motions that make up the total motion, where the shear complexity of the total motion prevents a complete parameterization based on experimental data only. On the other hand, the atomic detail provided by molecular dynamics simulation can, in principle, be used to separate the in- fluence of these multiple motions on the reorientational dynamics of individual bonds. Therefore, we develop a frame analysis, which allows the careful separation of multiple motions based on the MD trajectory. The separated motions may be analyzed and described with a few parameters. Using detector analysis [1,2], we then com- pare results from the simulated frame analysis to experimental re- INTRODUCTION sults, and refine parameters describing the separated motions us- SPONSORS ing experimental results. We apply the proposed to method to POPC PROGRAM bilayers, a critical biological interface, resulting in a dynamic land- Dynamic landscape of POPC. The landscape is derived based PROGRAM — BY DAY scape: a comprehensive description of motion in POPC molecules, on a combination of NMR and MD data. Motional amplitude is TOPICS for which we obtain the generalized amplitude of reorientational plotted as a function of correlation time and position. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] A. A. Smith, M. Ernst, B.H. Meier, J. Chem. Phys. 2018, 148, 045104. [2] A. A. Smith, M. Enrst, S. Riniker, B.H. Meier, Angew. Chem. Int. Ed. 2019, 131, 9483-9488. PT046 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULAR DYNAMICS 123 JENNIFER H. TOMLINSON1,2, ANTIBIOTIC RESISTANCE DRIVEN BY CHANGES IN ARNOUT P KALVERDA1,2, CONFORMATIONAL DYNAMICS ANTONIO N. CALABRESE1,2 1School of Molecualr and Cellular Biology, Fusidic acid (FA) is one of few remaining antibiotics that can be used to treat MRSA but resistance has increased University of Leeds, Leeds, UK 2 rapidly in recent years due to the expression of the FusB family of proteins. FA inhibits bacterial protein synthesis Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK by binding to Elongation Factor G (EF-G) when it is bound to the ribosome and preventing its release. FusB binds to EF-G and promotes the dissociation of these stalled complexes. The solution structure of the EF-G :FusB com-C3 plex identified both conformational changes and a significant change in the conformational flexibility of EF-G in response to FusB binding [1]. In this study, methyl relaxation dispersion NMR experiments have been used to characterize the difference in EF-G conformational flexibility in the apo and FusB bound states showing a widespread change in the conformational flexibility of domain 3, reflecting a significant increase in a minor state. HD exchange mass spectrometry suggests this results from increased domain 3 disorder upon FusB binding. Mutations disrupt-ing the effect of FusB binding on conformational flexibility but not affecting structural changes reduce the ability of FusB to confer FA resistance, showing the changes in dynamics are important in the resistance mechanism. These experiments show that FusB confers FA resistance though a significant change in the dynamics of domain 3 of EF- G, spreading throughout the domain and driving release of EF-G from the ribosome in a novel antibiotic resistance mechanism [2]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] JH Tomlinson, G.S. Thompson, AP. Kalverda, A. Zhuravleva, A.J.O’Neill Sci. Rep. 2016, 6, 19524. [2] J.H. Tomlinson, A.P. Kalverda, A.N. Calabrese, AUTHOR INDEX Proc. Natl. Acad. Sci. USA. 2020, 117, 25523-25531. PT047 PARALLEL SESSIONS / PROMOTED TALK / BIOMOLECULAR DYNAMICS 124 ALDO R. CAMACHO-ZARCO1, MOLECULAR BASIS OF HOST-ADAPTATION SISSY KALAYIL2, INTERACTIONS BETWEEN INFLUENZA VIRUS DAMIEN MAURIN1, NICOLA SALVI1, POLYMERASE PB2 SUBUNIT AND ANP32A ELISE DELAFORGE1, SIGRID MILLES1, Influenza A virus (IAV) is responsible for 3–5 million severe cases every year, resulting in 250-500,000 deaths. Most MALENE R JENSEN1, influenza strains evolve exclusively in the large reservoir of water birds, but some highly pathogenic avian strains DARREN J HART1, ( e.g. , H5N1, H5N8 and H7N9) can infect humans with lethal consequences (up to 60% mortality) and are potential STEPHEN CUSACK2, pandemic threats for humanity if they develop human-to-human transmissibility [1]. MARTIN BLACKLEDGE1 Avian influenza polymerase undergoes host adaptation in order to efficiently replicate in human cells [2]. Adap-1Institut de Biologie Structurale; Université tive mutants are localized on the C-terminal (627-NLS) domains of the PB2 subunit. In particular, mutation of PB2 Grenoble Alpes, CNRS, CEA, IBS, Grenoble, residue 627 from E to K rescues polymerase activity in mammalian cells. A host transcription regulator ANP32A, France 2European Molecular Biology Laboratory - comprising a long C-terminal intrinsically disordered domain (IDD), is responsible for this adaptation [3]. Human EMBL, Grenoble, France ANP32A IDD lacks a 33 residue insertion compared to avian ANP32A, and this deletion restricts avian influenza polymerase activity. We used NMR to determine conformational ensembles of E627 and K627 forms of 627-NLS of PB2 in complex with avian and human ANP32A [4]. Human ANP32A IDD transiently binds to the 627 domain, exploiting multivalency to maximize affinity. E627 interrupts the polyvalency of the interaction, an effect compensated by an avian-unique motif in the IDD. The observed binding mode is maintained in the context of heterotrimeric influenza polymerase, INTRODUCTION placing ANP32A in the immediate vicinity of known host-adaptive PB2 mutants. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES References: [1] H. Nicholls, PLoS Biol. 2006, 4, e50. [2] J.W. Almond, Nature. 1977, 270, 617-618. [3] J.S. Long, E.S. Giotis, et al, Nature. 2016, 529, 101-104. [4] PLENARY LECTURES A.R. Camacho-Zarco, S. Kalayil, D. Maurin , et al. Nat Commun. 2020, 11, 3656. TUTORIAL LECTURE Acknowledgements: This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie INVITED AND PROMOTED LECTURES Skłodowska-Curie grant agreement no. 796490 (AvInfluenza) and HFSP postdoctoral HFSP fellowship LT001544/2017 ‘Molecular basis of avian influenza polymerase adaptation to human hosts’. This work used the platforms of the Grenoble Instruct-ERIC centre (ISBG; UMS 3518 CNRS-CEA-UGA-EMBL) within POSTERS the Grenoble Partnership for Structural Biology (PSB), supported by FRISBI (ANR-10-INBS- 05-02) and GRAL, financed within the University Grenoble Alpes ADVERTISERS graduate school (Ecoles Universitaires de Recherche) CBH-EUR-GS (ANR-17-EURE-0003). IBS acknowledges integration into the Interdisciplinary Research AUTHOR INDEX Institute of Grenoble (IRIG CEA). IN041 PARALLEL SESSIONS / INVITED SPEAKER / SMALL MOLECULES II 125 PREDRAG NOVAK INTERACTIONS OF MACROLIDE ANTIBIOTICS AND University of Zagreb, Faculty MACROZONES UNDER THE SCOPE OF NMR of Science, Department of Chemistry, Zagreb, Croatia Macrolide antibiotics, such as azithromycin have been in clinical use for over 60 years, mostly due to their high efficacy, safety and favourable pharmacokinetics [1]. Macrolides bind to the 23S rRNA of the 50S subunit, near or at the peptidyl transferase center (PTC), block the nascent peptides exit tunnel and thus inhibit the synthesis of bacterial proteins. Multi-drug resistant microbial pathogens compromise their use as effective antimicrobials and poses seri-ous treats to human health worldwide which demand novel and more potent antimicrobial agents to be discovered. An effective approach to overcoming this problem is to understand the principles of how these drugs interact with their biological targets. Macrozones belong to a novel class of azithromycin-thiosemicarbazone conjugates that exhibit very good antibacterial activities against both susceptible and resistant bacterial strains [2]. In this talk results of interaction studies of macrolide antibiotics and some selected macrozones with their biological targets will be presented [2]. A combination of NMR experiments such as transferred nuclear Overhauser effect spectroscopy (trNOESY), saturation transfer difference (STD), diffusion and solvent paramagnetic relaxation experiments (PRE) and molecular modelling has been employed to characterize binding epitopes and asses bound conformations [4,5]. The data so obtained can serve as a basis for design of novel compounds with an improved biological profile. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] B. Arsic, P. Novak, M.G. Rimoli, J. Barber, G. Kragol, F. Sodano, Macrolides. Properties, Synthesys and Applications. Berlin: De Gruyter; 2018. [2] I. Grgičević, I. Mikulandra, M. Bukvić, M. Banjanac, I. Habinovec, B. Bertoša, P. Novak , Int. J. Antimicrob. Agents. 2020, 56, 106147. [3] T. Jednačak, I. Mikulandra, INVITED AND PROMOTED LECTURES P. Novak , Int. J. Mol. Sci. 2020, 21, 7799. [4] S. Kosol, E. Schrank, M. Bukvić Krajačić, G. E. Wagner, H. Meyer, C. Göbl, G.N. Rechberger, K. Zangger, P. Novak, J. POSTERS Med. Chem. 2012, 55, 5632–5636. [5] P. Novak, J. Barber, A. Čikoš, B. Arsić, J. Plavec, G. Lazarevski, P. Tepeš and N. Košutić-Hulita, Bioorg. Med. Chem. 2009, 17, ADVERTISERS 5857-5867. AUTHOR INDEX Acknowledgements: This work has been financially supported by the Croatian Science Foundation (project The Macrozones, IP-2018-01-8098). IN042 PARALLEL SESSIONS / INVITED SPEAKER / SMALL MOLECULES II 126 JONATHAN R. J. YONG,1 IMPROVING SENSITIVITY AND VERSATILITY IN NMR ALEXANDAR L. HANSEN,2 SUPERSEQUENCES ERIKS KUPCE,3 TIM D. W. CLARIDGE1 The structure characterisation of small molecules by NMR spectroscopy routinely employs well established 2D 1Chemistry Research Laboratory, homonuclear and heteronuclear correlation experiments such as COSY, TOCSY, HSQC, HMBC, NOESY, ROESY and Department of Chemistry, University of their many variants. In the arena of synthetic chemistry where many novel compounds require rapid structure Oxford, UK 2Campus Chemical Instrument Center, verification, or in metabolomics where multiple biomarkers require identification, there is significant interest in The Ohio State University, USA developing experimental methods that allow more rapid data collection to enable such strucutral identification. 3Bruker UK Ltd., U.K. NOAH (NMR by Ordered Acquisition using 1H-detection) represents one approach to fast data collection that re- cords multiple 2D data sets nested as specfically engineered aquisition modules within a single “supersequence”. [1,2] This approach requires only a single recovery delay for each series of nested modules, and so allows for significantly reduced data collection times. In the work presented here, the sensitivity and versatility of NOAH supersequences are improved by modification of sensitivity-enhanced HSQC and HSQC-TOCSY experiments, adding further versatility for 13C and 15N modules. [3] Importantly, these heteronuclear modules have been specifically tailored to preserve the magnetisation required for subsequent acquisition of other heteronuclear or homonuclear modules in a supersequence. In addition, we present protocols for optimally combining HSQC and HSQC-TOCSY/COSY elements within the same supersequenc- es, and further demonstrate that the associated time savings derived from NOAH can translate to increased detection sensitivity per unit time. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] E. Kupce and T. D. W. Claridge, Angew. Chem. Int. Ed., 2017, 56, 11779-11783. [2] E. Kupce and T. D. W. Claridge, Chem. Comm., 2018, 54, 7139-7142. AUTHOR INDEX [3] J. R. J. Yong, A. L. Hansen, E. Kupce., T. D. W. Claridge, 2021, submitted. PT048 PARALLEL SESSIONS / PROMOTED TALK / SMALL MOLECULES II 127 MATTHIAS BRAUSER, DETECTION AND VERIFICATION OF A KEY TIM HEYMANN, INTERMEDIATE IN AN ENANTIOSELECTIVE CHRISTINA MARIE THIELE Clemens-Schöpf-Institute for Organic PEPTIDE CATALYZED ACYLATION REACTION Chemistry and Biochemistry, Technical University of Darmstadt, Darmstadt, The selective acyl transfer onto alcohols is done in nature by acyltransferases. They are used in many important Germany signaling pathways, for example in plants for the synthesis of volatile esters while fruits are ripening, or in animals for the synthesis of the neurotransmitter acetylcholin. [1,2] Trying to reproduce the selectivity of this important reaction and to make it usable by chemists, Miller et al. introduced smaller peptide catalysts. Bearing a π-methylhistidine moiety these peptides were able to transfer acyl groups from acetic anhydride onto racemic alcohols while producing enantiomeric excess in the acylated product. [3] Schreiner and coworkers were able to build upon this success by also incorporating a π-methylhistidine moiety into a less flexible tetrapeptide. This peptide is able to reach enantiomeric excess values of > 99% for the acylation of racemic trans-cycloalkane-1,2-diols in toluene. [4] We herein report the detection of a key intermediate imidazolium-ion of the tetrapeptide, postulated to transfer the acyl-group onto the diols. Testing three different anhydrides in toluene and dichloromethane, we were able to identify imidazolium-ions for all of the anhydrides using ESI-HRMS. To further verify the existence of these ions in solution, HMBC- and selective NOESY-spectra were measured. Furthermore, a model reaction to test the prepared solution for the desired reactivity, was performed. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Y.-K. Yauk, E. J. F. Souleyre, A. J. Matich, X. Chen, M. Y. Wang, B. Plunkett, A. P. Dare, R. V. Espley, S. Tomes, D. Chagné, R. G. Atkinson, The Plant INVITED AND PROMOTED LECTURES Journal 2017, 91, 292-305. [2] D. Nachmansohn, A. L. Machado, Journal of Neurophysiology 1943, 6, 397-403. [3] S. J. Miller, G. T. Copeland, N. Papaioannou, POSTERS T. E. Horstmann, E. M. Ruel, Journal of the American Chemical Society 1998, 120, 1629-1630. [4] C. E. Müller, L. Wanka, K. Jewell, P. R. Schreiner, Angewandte ADVERTISERS Chemie International Edition 2008, 47, 6180-6183. AUTHOR INDEX Acknowledgements: We thank Peter Schreiner and Raffael C. Wende (Justus Liebig University of Giessen) for providing the tetrapeptide. PT049 PARALLEL SESSIONS / PROMOTED TALK / SMALL MOLECULES II 128 VICTOR RIBAY2, LED-NMR INVESTIGATION OF SPIRO-γ-LACTAM FORMATION MAUD VILLENEUVE1, DIDIER DURAND1, Spiro compounds are interesting building blocks in medicinal LED-NMR [3] is a powerfull method to investigate photochemical JÉRÔME-BENOÎT STARCK1, chemistry. These compounds have a three-dimensional structure reactions. Also, we followed the above described reaction using PHILIPPE HENNIG2, that allows them to occupy the space in a pocket of a targeted pro- this approach, with an already described LED setup [4] on a Bruker CHRISTOPHE PÉAN2 tein of interest. They are also interresting moieties for a fragment AVIII spectrometer, equipped with an inverse Prodigy® cryoprobe. 1 based approach of drug design (FBDD). The LED setup was constituted of a 420 nm LED light source for mi- Drug Design Small Molecules Unit, Institut de Recherches Servier, croscope illumination (Mic-LED-420Z, Prizmatix), fixed on a beam In 2020, Ryder et al. [1] described a photocatalyzed synthesis route Croissy-sur-Seine, France collimator coupled to a PMMA optic fiber (1000 mm core, 6 m of 2Drug Design Small Molecules Unit, for γ-spirolactam compounds, starting from the corresponding length). Analytical Department, Institut de primary amine. This synthetic approach was performed in the Recherches Servier, Suresnes, France presence of 4CzIPN [2] as the photocatalyst (PC), tetrabutylammo- We investigated the photoredox catalysis reaction with three cyclic nium azide as the HAT catalyst, one equivalent of methyl acrylate primary amines as starting materials, in CD CN. We followed, by 1D 3 (or 2-methoxyethyl acrylate) as the alkylating agent, without any and 2D NMR experiments, the quantitative a-tertiary amine N-func-protection of the amino group of the primary amine, at 425 nm in tionalisation, using methyl acrylate or 2-methoxyethyl acrylate as acetonitrile, and at room temperature. The corresponding γ-amino alkylating agent. Then, we determined the kinetic constants and ester was formed as intermediate of synthesis, and, after an intra-the reactional rate of the reaction for these three different amines, molecular reaction of elimination in presence of triethylamine in and highlighted the spontaneous ring formation of the lactam in methanol solution under reflux, the ring closure formation of the the reaction medium at room temperature, without additional tri- γ-lactam was then observed. ethylamine treatment. INTRODUCTION LED-NMR has proven to be a powerful method for photochemical SPONSORS reactions monitoring, and can replace LC-MS as analytical tool in PROGRAM the absence of chromophores on chemical species. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] A. S. H. Ryder, W. B. Cunningham, G. Ballantyne, T. Mules, A. G. Kinsella, J. Turner-Dore, C. M. Alder, L. J. Edwards, B. S. J. McKay, M. N. Grayson, A. J. Cresswell, Angew. Chem. ADVERTISERS Int. Ed. 2020, 59, 14986– 1499. [2] J. Luo, J. Zhang, ACS Catal. 2016, 6, 873–877. [3] P. Nitschke, N. Lokesh, R. M. Gschwind, Prog. Nucl. Magn. Reson. Spectrosc., 2019, 114–115, 86-134. [4] Y. Ji, D. AUTHOR INDEX A. DiRocco, C. M. Hong, M.K. Wismer, M. Reibarkh, Org. Lett. 2018, 20, 2156−2159. PT050 PARALLEL SESSIONS / PROMOTED TALK / SMALL MOLECULES II 129 ELENA PIERSANTI1, CHALLENGES IN THE DECOMPOSITION OF NMR AFEF CHERNI2, SPECTRA OF SMALL MOLECULE MIXTURES SANDRINE ANTHOINE2, CAROLINE CHAUX2, Despite the development of Nuclear Magnetic Resonance (NMR) methods to increase spectral resolution, the grow- LAETITIA SHINTU1, ing complexity of the samples leads to crowded spectra that compromise the analytical performances of this tech-MEHDI YEMLOUL1, nique [1]. The association of mathematical methods for signal processing with the methodological developments BRUNO TORRÉSANI2. in NMR is a promising alternative [2, 3]. In this context, this work is focused on the analysis of complex mixtures 1Aix Marseille, CNRS, Centrale by NMR with two aspects: the development and optimization of NMR pulse sequences and the application of Blind Marseille, iSM2, Marseille, France, 2 Source Separation (BSS) algorithms to NMR data. This source separation technique, originally used for disciplines Aix Marseille Univ, CNRS, Centrale Marseille, I2M, Marseille, France such as acoustics for audio signal processing, has shown its effectiveness for the demixing of 1D and 2D NMR spectra [4-7]. In this case, spectra decomposition is performed using correlations, essentially variations in concentrations, detected over a series of data sets, which allows the extraction of the pure spectra of the mixture constituents. Several types of samples such as synthetic mixtures of terpenes are used to evaluate the efficiency of algorithms for the extraction of the pure spectra. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] K. Kim, Y. H. Choi, R. Verpoorte, Trends Biotechnol. 2011, 29, 267-75. [2] C. Chaux, P.L. Combettes, J. C. Pesquetand, V.R.Wajs, Inverse Problems. 2007, 23, 1495 – 1518. [3] P. Comon, C. Jutten, Academic press. 2010. [4] D. Nuzillard, S. Bourg, J. M. Nuzillard, J. Magn. Reson. 1998, 133, 358-363. [5] I. Toumi, INVITED AND PROMOTED LECTURES S. Caldarelli, B. Torrésani, Prog. Nucl. Mag. Res. Sp. 2014, 81, 37 – 64. [6] I. Toumi, B. Torrésani, S. Caldarelli, Anal. Chem. 2013, 85, 11344 – 11351. [7] A. Cherni, POSTERS E. Piersanti, S. Anthoine, C. Chaux, L. Shintu, M. Yemloul, B. Torrésani, Faraday Discuss. 2019, 218, 459-480. ADVERTISERS Acknowledgements: This project has received funding from the Excellence Initiative of Aix-Marseille University – A*midex, a French “Investissement AUTHOR INDEX d’Avenir” program. IN043 PARALLEL SESSIONS / INVITED SPEAKER / FRONTIERS IN MAGNETIC RESONANCE 130 FEDOR JELEZKO NANOSCALE MAGNETIC RESONANCE ENABLED BY Ulm University, Ulm, Germany DIAMOND QUBITS A particularly interesting application of diamond based quantum sensing is the detection of nuclear magnetic resonance on nanometer scales, including the detection of individual nuclear spins or small ensembles of external nuclear spins. Single nitrogen vacancy (NV) color centers in diamond currently have sufficient sensitivity for detecting single external nuclear spins and resolve their position within a few angstroms. The ability to bring the sensor close to biomolecules by implantation of single NV centers and attachment of proteins to the surface of diamond enabled the first proof of principle demonstration of label-free detection of the signal from a single protein. Single-molecule nuclear magnetic resonance experiments open the way towards unraveling dynamics and structure of single biomolecules. However, for that purpose, NV magnetometers must reach spectral resolutions comparable to that of conventional solution state NMR. New techniques for this purpose will be discussed. We will also show first experiments towards hyperpolarisait of extrneal nuclear spins using shallow NV centers. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN044 PARALLEL SESSIONS / INVITED SPEAKER / FRONTIERS IN MAGNETIC RESONANCE 131 JÖRG WRACHTRUP PROBING MATERIALS WITH SINGLE SPIN QUANTUM University of Stuttgart, 3rd SENSORS Institute of Physics and Center for Applied Quantum Technology, Stuttgart, Germany Single spin probes are developing into versatile use to measure the properties of materials with high spatial resolution. Besides measuring diffusion through single spin detected NMR [1], specifically magnetic materials lend themselves as interesting objects to be investigated by single spin quantum probes. I will show how to measure the micro and nanomagnetic properties of two-dimensional material by measuring its local magnetic field [2]. To this end, a single spin system, like a single nitrogen vacancy centre, is scanned across a sample while at the same time measuring its electron spin resonance frequency. I will demonstrate that one can achieve a spatial resolution of a few 10 nm with a magnetic field sensitivity of below 1 mT. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] F. Shagieva et al. , arxiv:2105.07712. [2] Qi-Chao Sun et al. , Nature Comm. 2021, 12, 1989. IN045 PARALLEL SESSIONS / INVITED SPEAKER / FRONTIERS IN MAGNETIC RESONANCE 132 TJERK OOSTERKAMP MAGNETIC RESONANCE FORCE MICROSCOPY: Leiden Institute of Physics, TOWARDS SINGLE ELECTRON SPIN DETECTION AND Leiden University, Leiden, The Netherlands NANOMRI In this presentation, we will introduce new methods and their challenges in MRFM, where magnetic resonance is generating forces that are detected using Atomic Force Microscopy. We will discuss particularly how the challenges of working at temperatures below 1 Kelvin might be addressed. We present data in which we show that nuclear magnetic resonance of copper allows nanoscale imaging, and discuss how this translates to the feasibility of using Boltzmann polarization of protons for nanoscale imaging, even at 1 MHz. Finally, we present the challenges of Force measurements on magnetic resonance of single electrons. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN046 PARALLEL SESSIONS / INVITED SPEAKER / FRONTIERS IN MAGNETIC RESONANCE 133 LUANA DE CAMARGO1, HIGHLY COHERENT AND PROCESSABLE MATTEO BRIGANTI1,2, ORGANOMETALLIC SPIN QUBITS GIULIA SERRANO2, ANDEA LUIGI SORRENTINO2, Organometallic sandwich complexes of paramagnetic lanthanides ions are currently the focus of great interest be-LORENZO POGGINI2,3, cause of the observation that dysprosium(III) derivatives present magnetic hysteresis of molecular origin, i.e. Sin-MATTEO MANNINI2, gle-Molecule Magnet behavior, above liquid nitrogen temperature [1]. LORENZO SORACE2, FEDERICO TOTTI2, Sandwich complexes of early transition metal ions such as Ti and V carrying one unpaired electron are also well ENRICO SALVADORI4, known but their spin dynamics not equally investigated. In our earch for highly coherent and evaporable magnetic MARIO CHIESA4, molecules as potential molecular spin qubits, we identified in [CpTi(cot)] and [CpV(cht)] where Cp= η5-C H , cot= 5 5 η8- JAISA F. SOARES1, C H , and cht= H , promising candidates [2]. Pulsed EPR investigations have revealed that the coherence times 8 8 η7-C7 7 ROBERTA SESSOLI2 reach ca.35 ms at low temperature in frozen deuterated toluene, despite the molecules being hydroge-rich. Ab initio 1 calculations revealed that the low energy vibrational modes involving the rings rotation have a weak spin-phonon Dept. of Chemistry, Federal University of Parana, Curitiba, Brazil coupling, while hydrogen atoms are placed within the frozen sphere, thus not too detrimental for decoherence. 2Dept. of Chemistry University of Florence, Sesto Fiorentino, Italy Moreover these small neutral molecules can be evaporated and deposited on different substrates with a moderate 3 ICCOM CNR, Sesto Fiorentino, Italy hybridization with the underlying metal, whose extent depends on the metal substrate and on the adsorption ge- 4Dept. of Chemistry University of Turin, Turin, Italy ometry [3]. They appears very promising candidates for single spin addressing via Scanning Tunnel Microscopy and for tip functionalization with a spin probe [4]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] F.-S. Guo et al., Science 2018, 362, 1400. [2] L. C. de Camargo et al- Angew. Chem. Int. Ed. 2021, 60, 2588. [3] G. Serrano et al. in preparation. [4] INVITED AND PROMOTED LECTURES B. Verlhac, et al., Science 2019, 366, 623. POSTERS Acknowledgements: We thank the financial support by the QUANTERA project SUMO, the H2020-FET-OPEN project FATMOLS (GA 862893), and by the ADVERTISERS Brazilian CNPq (Project 308426/2016-9) and CAPES AUTHOR INDEX (PROEX and PrInt/CAPES-UFPR, Finance Code 001). PT051 PARALLEL SESSIONS / PROMOTED TALK / FRONTIERS IN MAGNETIC RESONANCE 134 PETR ŠTĚPÁNEK OPTICAL CONTRASTS IN NMR USING NUCLEAR NMR Research Unit, Faculty of MAGNETO-OPTIC EFFECTS Science, University of Oulu, Oulu, Finland In a typical NMR experiment, the information about the chemical structure of a molecule is encoded in the chemical shift, while the intensities of the NMR peaks carry quantitative information about the ratio of equivalent nuclei. In addition to classical NMR, the nuclear magnetization can also be detected using polarized light [1]. The measured signal in these so-called nuclear magneto-optic (NMO) effects originates from a complex interplay of the nuclear magnetic moments, electron density of the molecule, and the light beam. In contrast to NMR, the intensity of NMO signal qualitatively depends on the local chemical environment around the nucleus. This effect is called optical chemical shift [2] and offers additional possibility for advanced resolving of complex molecular structures. In this contribution I present our recent theoretical investigations on this topic [3]. In particular I discuss the NMO effect called nuclear spin-induced optical rotation (NSOR), which is a rotation of linearly polarized light by nuclear magnetization oriented parallel to the light beam. By investigating 1H and 13C NSOR spectra of a large set of hydrocarbons, patterns are predicted showing systematic relation between the intensity of NSOR and the local molecular structure. The signal intensities are grouped according to the chemical bonding near the nucleus and its neighbors, their positions within the molecule with respect to other nuclei, and local isomerism. This additional information can be used to distinguish the nuclei and help in resolving spectra of complex systems. Further experimental development of NSOR technique and combining it with existing NMR methods could provide INTRODUCTION exciting opportunities for completely new kind of multidimensional experiments for higher resolution spectrosco- SPONSORS py and measurements of new molecular properties. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] I. M. Savukov, S.-K. Lee, M. V. Romalis, Nature 2006, 442, 1021-1024. [2] S. Ikäläinen, M. V. Romalis, P. Lantto, J. Vaara, Phys. Rev. Lett. 2010, 105, POSTERS 153001. [3] P. Štěpánek, Phys. Chem. Chem. Phys. 2020, 22, 22195-22206. ADVERTISERS Acknowledgements: The author is grateful for the financial support from Academy of Finland (Grant 316180), the Kvantum institute (University of Oulu), and AUTHOR INDEX grants of computer capacity from the Finnish Grid and Cloud Infrastructure (persistent identifier urn:nbn:fi:research-infras-2016072533). PT052 PARALLEL SESSIONS / PROMOTED TALK / FRONTIERS IN MAGNETIC RESONANCE 135 DAVID E KORENCHAN, SINGLET-STATE 31P NMR SPECTROSCOPY BETWEEN CHEMICALLY ALEXEJ JERSCHOW EQUIVALENT NUCLEI Deparment of Chemistry, New York University, New York, NY, USA The discovery of long-lived nuclear magnetic resonance (NMR) sin- constant 2J of 11.6 Hz.The differential couplings between 1H and 31P HH glet states has been an exciting area of scientific inquiry over the nuclei enable access to a 31P singlet state. We measured spin-lattice past two decades. The attractive possibility to store phase informa- (R ) and singlet (R ) relaxation rates at 9.4 T at various temperatures 1 s tion or population differences over long periods of time relative to using inversion-recovery and spin level induced crossing (SLIC) [4] other NMR timescales has garnered interest for using singlets to sequences, respectively. Surprisingly, R exceeded R considerably at s 1 study slow molecular rearrangements, diffusion, and hyperpolar- all temperatures measured, contrary to expected results. In order ization storage [1]. NMR singlets have even been postulated to play to elucidate which mechanisms dominate singlet relaxation, we a role in biological processes, particularly between 31P nuclei in bio-conducted molecular dynamics (MD) simulations with AMBER and molecules [2]. We report the generation and lifetime measurement calculated chemical shift anisotropy (CSA) tensors with Gaussian. of a 31P singlet for a chemically equivalent case in tetrabenzyl pyro- We found that the unexpectedly short relaxation rates could be ex- phosphate (TBPP). plained by the very large CSA present between 31P nuclei in TBPP. The two CSA tensors for each phosphate are highly anticorrelated The parameters of the unusually complex 31P and 1H spectra, caused and at a relatively constant angle with one another, leading to en- by magnetic inequivalence, were extracted by comparing the ex- hanced relaxation. perimental results with simulations using the Spinach MATLAB package [3]. We modeled the compound as two 31P nuclei, each In conclusion, we demonstrate the existence of a singlet state be-J-coupling with 4 1H nuclei that are 3 bonds away. Excellent agree- tween magnetically inequivalent 31P nuclei. We observe a very short INTRODUCTION ment with experimental results was further found by using the fol- 31P singlet lifetime in TBPP dominated by large uncorrelated CSA SPONSORS lowing J-coupling parameters: 2J = 16.4 Hz and 3J = 8.4 Hz. We interactions. It is likely that the singlet lifetimes are significantly PP PH PROGRAM also discovered by spectral simulation that the 1H nuclei in each pair longer at low magnetic fields. Future work will focus on 31P singlets PROGRAM — BY DAY of methylene protons are chemically inequivalent, having a chemi- in other compounds, including biological species. TOPICS cal shift difference of 0.0089 ppm (3.6 Hz at 9.4 T) and a J-coupling PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] M. Levitt, J. Magn. Reson. 2019, 306, 69-74. [2] M. P. A. Fisher, Ann. Phys. 2015, 362, 593-602. [3] H. J. Hogben, M. Krzystyniak, G. T. P. Charnock, P. J. Hore, I. Kuprov, J. Magn. ADVERTISERS Reson. 2011, 208, 179-194. [4] S. J. DeVience, R. L. Walsworth, M. S. Rosen, Phys. Rev. Lett. 2013, 111, 173002. AUTHOR INDEX Acknowledgements: We acknowledge funding though an award of the U.S. National Science Foundation, award no. CHE 1710046, and an award by the Heising Simons Foundation. PT053 PARALLEL SESSIONS / PROMOTED TALK / FRONTIERS IN MAGNETIC RESONANCE 136 BRUNO SIMÕES DE ALMEIDA, RESONANCES IN 1H MAS SPECTRA ARE NOT CENTERED PINELOPI MOUTZOURI, AT THE ISOTROPIC CHEMICAL SHIFT GABRIELE STEVANATO, LYNDON EMSLEY. In solid-state NMR, the homonuclear dipolar coupling is the internal spin interaction that has the greatest con-Institut des Sciences et Ingénierie Chimiques, tribution to the broadening of the 1H spectral lines. Even at the fastest magic-angle spinning rates available today Ecole Polytechnique Fédérale de Lausanne [100 kHz-150 kHz], the linewidths can extend over hundreds of Hertz, affecting 1H resolution. Understanding and (EPFL), Lausanne, Switzerland minimizing this contribution could lead to rich structural information for organic solids. In the past, systems of two and three spins were studied with Average Hamiltonian Theory [1] (AHT) to second order and Floquet Theory [2-3]. Here we study two and three inequivalent spin systems (I=½) with AHT, in the fast-spinning regime, and we develop analytical expressions of the average Hamiltonian to 3rd order. The results show that the full expression of the 3rd order average Hamiltonian, without secular approximation or truncation to second-order, agrees the best with full numerical calculations. We find that the effect on the NMR spectrum of the different Hamiltonian terms, is to produce both MAS rate dependent residual shifts and splittings in the three-spin case. Powder lineshapes are also analyzed, and it is found that the anisotropic residual shift does not have zero average, indicating that the lineshape is broadened and shifted from the isotropic position, and we confirm this with experimental observations for 1H MAS spectra in molecular solids [4-5]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] M. M. Maricq, J. S. Waugh, J. Chem. Phys. 1979, 70, 3300-3316. [2] M. H. Levitt, D. P. Raleigh, F. Creuzet, R. G. Griffin, J Chem Phys 1990, 92, 6347-6364. [3] Nakai, C. A. McDowell, J Chem Phys POSTERS 1992, 96, 3452-3466. [4] P. Moutzouri, F.M. Paruzzo, B. S. de Almeida, G. Stevanato, L. Emsley, Angewandte Chemie 2020, 59(15), 6235-6238 [5] P. Moutzouri, B. Simoes de Almeida, L. Emsley, J Magn. Reson. 2020, 321, 106856. ADVERTISERS AUTHOR INDEX Acknowledgements: EPFL- Laboratory of Magnetic Resonance, Swiss National Science Foundation Grant No. 200020_178860. IN047 PARALLEL SESSIONS / INVITED SPEAKER / BIOSOLIDS 137 MARKUS WEINGARTH UNDERSTANDING ANTIBIOTICS WITH NMR Spectroscopy, Department SOLID-STATE NMR of Chemistry, Utrecht University, Utrecht, The Netherlands Antibiotics that use novel mechanisms are needed to resolve the antimicrobial resistance crisis. Promising tem- plates could be antibiotics that target Lipid II, known as the Achilles’ heel of bacteria, at a conserved and immutable pyrophosphate group. Such antibiotics, like plectasin, teixobactin, or malacidin [1-3] would kill the most refractory pathogens without causing resistance. However, due to the challenge of studying small antibiotic-target complexes in membranes, the structural correlates of the relevant binding modes are virtually unknown. Here, using solid-state NMR in combination with microscopy, we report on the physiologically relevant binding modes of several Lipid II-binding antibiotics [4-6]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Schneider et al. , Science 2010; [2] Ling et al. , Nature 2015; [3] Hover et al. , Nature Microbiology 2018; [4] Medeiros - AUTHOR INDEX Silva et al. , Nature Communications 2018; [5] Shukla et al., Nature Communications 2020; [6] Shukla et al. , submitted. IN048 PARALLEL SESSIONS / INVITED SPEAKER / BIOSOLIDS 138 CLEMENS GLAUBITZ COMBINING PHOTOCHEMISTRY AND SOLID-STATE Biophysical Chemistry and NMR FOR MEMBRANE PROTEIN RESEARCH Centre for Biomolecular Magnetic Resonance (BMRZ), Goethe University Frankfurt, The use of solid-state NMR for resolving the mode of action of photochemical tools in lipid bilayers and for mecha-Frankfurt am Main, Germany nistic studies on photoreceptors is demonstrated and discussed. First, it is shown by MAS-NOESY and R-PDLF experiments how photoswitchable lipids within lipid bilayers alter the general membrane properties [1]. The light-induced trans-cis isomerisation of these AzoPCs causes localized disorder in the membrane. The embedded integral E. coli membrane protein diacylglcerol kinase (DgkA) serves as a model to probe how these effects can influence protein structure and activity. The known resonance assignment of DgkA [2] enabled to record a series of 2D NCA spectra after illumination from which site-resolved, light-induced effects could be visualized. Protein activity has been assessed by real-time 31P MAS NMR. Our data reveal in detail how photositchable lipids alter bilayer proerties and how these affect embedded membrane proteins. We also demonstrate how the enzymatic reaction catalyzed by DgkA can be triggeded by light under MAS NMR conditions [3]. While these experiments rely on high fields, extensive labelling and non-frozen samples, other experimental re- quirements apply when targetting photoreceptor intemediate states. Here, DNP-enhanced solid-state NMR in com- bination with light-induced cryo-trapping has been proven as an indispensable tool, which will be briefly illustrated by two recent examples. In case of channelrhodopsin-2 long-range double quantum recoupling sequences (SR26) and systematic cryo-trapping procedures enabled to differentiate between competing photocycle models [4], while INTRODUCTION in case of the sodium pump KR2, the chromophore configuration could be identified during the sodium transfer SPONSORS step [5]. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] M. Doroudgar, J. Moorstein, J. Becker.Baldus, D. Trauner, C. Glaubitz, J. Am. Chem. Soc. 2021, accepted. [2] K. Mobius, S. Kazemi, P. Guntert, A. Jakob, A. Heckel, J. Becker-Baldus, C. Glaubitz, Sci. Rep. 2019, 9, 3995. [3] J. de Mos, A. Jakob, J. Becker-Baldus, A. Heckel, C. Glaubitz. Chem. Eur. J. 2020, INVITED AND PROMOTED LECTURES 26, 6789. [4] J. Becker-Baldus, A. Leeder, L. J. Brown, R. C. D. Brown, C. Bamann, C. Glaubitz. Angew. Chem. Int. Ed. Engl. 2021, epub ahead of print. [5] O. POSTERS Jakdetchai, P. Eberhardt, M. Asido, J. Kaur, C. N. Kriebel, J. Mao, A. J. Leeder, L. J. Brown, R. C. D. Brown, J. Becker-Baldus, C. Bamann, J. Wachtveitl, C. Glaubitz. ADVERTISERS Si. Adv. 2021, 7, eabf4213. AUTHOR INDEX Acknowledgements: Deutsche Forschungsgemeinschaft, Graduate School CLIC. PT054 PARALLEL SESSIONS / PROMOTED TALK / BIOSOLIDS 139 CHI L. L. PHAM1, NECROSOME ASSEMBLY STUDIED BY SOLUTION AND MARGARET SUNDE1, SOLID-STATE NMR MIGUEL MOMPEÁN2 1Faculty of Medicine and Health Receptor-Interacting Protein Kinase 3 (RIPK3) is the key effector that activates the mixed lineage kinase domain-like and Sydney Nano, University of (MLKL) protein to execute necroptosis, a type of programmed cell death distinct from apoptosis [1]. Necroptosis Sydney, Sydney, Australia 2“Rocasolano” Institute of activation requires the formation of intermediate signaling complexes named necrosomes. RIPK3 a common com-Physical Chemistry, Spanish ponent in all distinct necrosome complexes, whose assembly is mediated by a conserved region named RIP Ho- National Research Council, motypic Interaction Motif (RHIM), with sequences I(V)QI(V/L/C)G. Of the different necrosomes, the RIPK1-RIPK3 Madrid, Spain complex is regarded as the canonical necrosome, and TRIF-RIPK3 and ZBP1-RIPK3 are other two non-canonical ne- crosomes, all leading to the activation of MLKL [2, 3]. All these complexes are amyloidal assemblies, and solid-state NMR studies on the RIPK1-RIPK3 canonical necrosome core revealed a 1:1 hetero-amyloid structure [4], changing our vision that amyloids are homo-polymers building on copies of a same type of protein. Density Functional Theory (DFT) calculations corroborated that 1:1 co-assembly is preferred over self-assembly. While ongoing solid-state NMR studies will reveal the key differences among distinct necrosomes, solution NMR has the unique potential to inform the conformational changes and interactions in free and amyloid-bound monomers to unveil the molecular determinants that drive homo- versus hetero-amyloid formation. Recent improvements in preparing, handling and stabilizing RHIM proteins are enabling for the first time characterizing of the monomeric form of RIPK3 and its oligomerization pathways in solution [5]. Analysis of chemical shifts and 15N relaxation measurements illustrate a chiefly disordered domain with key residues around the RHIM region showing slowed motion in the µs-ms times- INTRODUCTION cales, in agreement with the amyloid core mapped by both solid-state NMR and 15N-DEST (Dark-state Exchange SPONSORS Saturation Transfer) experiments that we are using to characterize the interaction sites in the assembled state. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] W. Sun, X. Wang, Trends Biochem. Sci. 2014, 39, 587-593. [2] W.J. Kaiser, et al., J. Biol. Chem. 2013, 288, 31268-31279. [3] M. Rebsamen, et al., INVITED AND PROMOTED LECTURES EMBO Rep. 2009, 10, 916-922. [4] M. Mompeán, et al., Cell. 2018, 173, 1244-1253. [5] C.L.L. Pham, et al., EMBO Rep. 2019, 20, e46518. POSTERS Acknowledgements: MM is a Ramón y Cajal Fellow of the Spanish AEI-Ministry of Science and Innovation (RYC2019-026574-I) and a “La Caixa” Foundation ADVERTISERS Junior Leader Fellow (LCR/bq/pr19/11700003). NMR experiments were performed in the “Manuel Rico” NMR laboratory (LMR) of the Spanish National AUTHOR INDEX Research Council (CSIC), a node of the Spanish Large-Scale National Facility (ICTS-R-LRB). PT055 PARALLEL SESSIONS / PROMOTED TALK / BIOSOLIDS 140 CLARA NASSRIN KRIEBEL1, FASTER MAS NMR DATA ACQUISITION AND JOHANNA BECKER-BALDUS1, FUNCTIONAL ANALYSIS OF THE LIGHT-DRIVEN ALIA HASSAN2, JOCHEM STRUPPE3, SODIUM PUMP KR2 CLEMENS GLAUBITZ1 1 Solid-state NMR (ssNMR) is a powerful technique to study membrane protein function, dynamics and structure in Goethe University, Institute for Biophysical Chemistry & Center for Biomolecular a native-like environment. High sensitivity is required for a complete resonance assignment based on highly re- Magnetic Resonance (BMRZ), Frankfurt am solved 3D- and 4D- heteronuclear 13C, 15N correlation spectra recorded at high magnetic fields. Here we combine sev-Main, Germany 2 eral techniques to improve sensitivity which enabled us to assign the first known light driven sodium pump Kroki-Bruker Switzerland AG, Fällanden, Switzerland nobacter eikastus rhodopsin 2 (KR2) [1-3], a hepta-helical membrane embedded protein. Paramagnetic Gd3+-doping 3Bruker Biospin Corporation, Billerica, MA, [4] allowed a 2,5-times faster acquisition using an E-free MAS probe optimized for a higher duty cycle. Double-CP USA optimum control pulse sequences [5] enabled 4-fold faster data recording. Non-uniform sampling shortened ex- perimental times 2-times [6]. BioSolids Cryoprobe additionally allowed a seven times faster data acquisition [7]. As a result, 3D and 4D spectra could be recorded in 5 and 12 days, respectively. The KR2 assignment covers more than 80% including numerous functional important residues which are mainly involved in the ion selectivity and pump-ing mechanism such as D116 and Q123 and allows the NMR analysis of functional KR2 mutants with a promising future application as optogenetic tools [8]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] Gushchin I. et al., Nat. Struct. Mol. Biol. 2015, 22(5), 390-395. [2] Inoue K. et al., Nat. Commun. 2013, 4, 1678. [3] Kato H. E. et al., Nature, 2015, ADVERTISERS 521(7550), 48-53. [4] Ullrich, S. et al., J. Biomol. NMR 2014, 58, 27–35. [5] Tošner Z. et al., Angew. Chem. Int. Ed. 2018, 57, 14514-14518. [6] Lin, E.C. et al., J. Magn. AUTHOR INDEX Reson. 2013, 237, 40-48 [7] Hassan, A. et al., J. Magn. Reson. 2020, 311(106680). [8] Zhang F. et al., Cell. 2011, 147(7), 1446-1457. PT056 PARALLEL SESSIONS / PROMOTED TALK / BIOSOLIDS 141 XAVIER FALOURD1,2, HOW SOLID-STATE AND LOW-FIELD NMR CAN CONTRIBUTE TO AMANDINE LEROY1,4, THE SUPRAMOLECULAR ORGANIZATION ELUCIDATION OF THE VALÉRIE MÉCHIN3, FABIENNE GUILLON1, LIGNOCELLULOSIC BIOMASS DURING DECONSTRUCTION? GABRIEL PAËS4 AND LOÏC FOUCAT1,2 Over the last 15 years, research efforts have been dedicated to iden- wall composition with a loss of hemicelluloses and of ferulic acid 1 tifying the factors that influence lignocellulosic biomass (LB) en- cross-linking, associated with a lignin enrichment. Biopolymères Interactions Assemblages (BIA) laboratory, zymatic deconstruction to develop alternatives to fossil carbon To characterize water dynamics, low-field T measurement were INRAE, Nantes, France, resources. Accessibility of LB to enzymes is an important proper- 2 2INRAE, BIBS Facility, Nantes, carried out. An inverse Laplace transformation was applied to con- ty involved in the recalcitrance in LB deconstruction related to the France, vert the relaxation signal into a continuous distribution of T . At the 3Institut Jean-Pierre Bourgin (IJPB), composition of the LB (hemicelluloses, lignin, hydroxycinnamic ac- 2 hydration level used in solid state experiments (~20% w/w), HWP INRAE-AgroParisTech, CNRS, ids content…) and dependent on such structural factors as chemical Université Paris-Saclay, Versailles, induced a decrease of the T peak width, indicating an homogeni- France, structure and interactions of cell wall polymers, cell wall porosity... 2 zation of water environments. In the enzymatic hydrolysis humid- 4Université de Reims Champagne This study focused on the hot water pretreatment (HWP) of maize Ardenne, INRAE, FARE, UMR A614, ity conditions (~80% w/w), a modulation of the structural domains internodes (180°C applied during 20 or 40 min) that induces a reor- Reims, France. due to HWP was observed, which can be associated with more water ganization of the 3D network. in pore sizes between 5 and 15 nm. This increase in meso-porosi- The impact of HWP on cellulose supramolecular organization was ty should favour the accessibility of enzymes and their catalytic investigated by 13C CPMAS. An increase of cellulose crystallinity activity. concomitant with an increase of agregate dimension was observed. This study underlines that the loss and changes in polymers struc- INTRODUCTION Changes in the lignin structure, in relation to the decrease of β-O-4 tural features induced by HWP leads to a reorganization of the cell SPONSORS bonds, were evidenced. To supplement these findings, polarization walls. This results in a water redistribution improving the accessi- PROGRAM transfer kinetics was studied. The dynamical parameters T H and 1r bility of cellulases and, consequently, the hydrolysis yield [2]. The PROGRAM — BY DAY T were thus determined using a two proton-reservoir model [1]. HH results obtained by the combined use of solid-state and low-field TOPICS HWP induced an increase of T H and T values associated with an 1r HH NMR modalities, associated with those obtained by chemical ana- PRIZE LECTURES increase of the molecular order and with the density of water mol- lyzes, were synthesized in a schematic representation illustrating PLENARY LECTURES ecules in the macromolecular assembly, respectively. In addition, the effect of HWP on the macromolecular assembly structure. TUTORIAL LECTURE the chemical characterization showed that HWP modified the cell INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] W. Kolodziejski, J. Klinowski, Chem. Rev.. 2002, 102, 613-628. [2] A. Leroy, X. Falourd, L. Foucat, V. Méchin, F. Guillon, G. Paës, Biotechnology for Biofuels. 2021, under revision. AUTHOR INDEX Acknowledgements: members of UMR FARE and UR BIA staff. Funding of Amandine Leroy PhD was provided by Région Grand-Est/Grand Reims. IN049 PARALLEL SESSIONS / INVITED SPEAKER / METABOLOMICS 142 DANIEL RAFTERY, QUANTITATIVE METABOLOMICS METHODS G.A. NAGANA GOWDA DEVELOPMENT USING NMR Northwest Metabolomics Research Center, One of the ultimate goals of the field of metabolomics is a fully quantitative analysis of highly complex biological Department of Anesthesiology and Pain Medicine, University of samples. Ideally, platforms that provide broad metabolome coverage allow the opportunity for deep insights into Washington, Seattle, WA, USA biological problems, while excellent quantitation ensures good data quality, opportunities for précising metabolic modeling, and allows an ability to compare across studies. However, these results can be difficult to achieve on a routine basis because the highly complex sample matrix often precludes reliable measurements of many metabolites and complicates quantitation efforts. NMR spectroscopy has much to offer the field of metabolomics due to its exquisite quantitative capabilities and reproducibility, especially in the area of new methods development. Recently, we have investigated the use of new reference compounds for improved metabolite quantitation. The common NMR reference compounds that work well for simple solutions or mixtures produce inaccurate results when used to quantitate blood metabolites. Our approach takes advantage of a simple protein precipitation procedure that allows the absolute quantitation of over 80 metabolites using a single standard compound. These metabolites, including even some at sub-micromolar concentrations, span a broad range of classes and pathways, including organic and amino acids, as well as energy metabolites and co-enzymes. We show that this approach is more robust than current approaches. We have also shown that this quantitative NMR approach is also useful for calibrating quantitative mass spectrometry measurements, without the use of internal or external metabolite standards and has led to some unusual findings about the stability of well-known metabolites. INTRODUCTION SPONSORS We are also very interested in understanding why metabolite levels can change from collection site to collection PROGRAM site. Some of these effects are likely caused by differences in sample treatment procedures and we are in the process PROGRAM — BY DAY of trying to model these with the goal of improving cross site validation studies. It may also be possible to develop TOPICS methods to mitigate the effects of sample mistreatment using either experimental or modeling approaches. Recent PRIZE LECTURES progress in our work in this emerging area will be discussed. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN050 PARALLEL SESSIONS / INVITED SPEAKER / METABOLOMICS 143 CAMILA CARDOZO1, UNCOVERING METABOLIC FEATURES OF CELL ABDERRAFEK EL HARRAS1, PLASTICITY AND CARCINOGENESIS MANHAL MILI2, ARIEH MOUSSAIEF3, Nuclear Magnetic Resonance (NMR) is a powerful platform for molecular phenotyping of biological systems and HIND HAFSI1, its highly versatile nature opens up a range of analytical strategies for metabolomic phenotyping of biological sys-EMILIE MONTELLIER1, tems. Here, we illustrate how NMR metabolic fingerprints of intra- and extra-cellular material provide a unique PIERRE HAINAUT1, experimental platform for cellular biology. We show how solution and HR-MAS 1H NMR investigations can provide BÉNÉDICTE complementary insights into the metabolism of pluripotent stem cells, and identify the early metabolic changes ELENA-HERRMANN1 associated with cell differentiation. 1Univ Grenoble Alpes, CNRS, INSERM, Institute for Advanced Biosciences, In a second part, we present a metabolomics investigation of TP53 mutations in primary human cell lines. TP53 is Grenoble, France the most mutated gene in cancer, with about 50% of cancers harboring a single missense mutation. Over the past 2Univ Lyon, Institute for Analytical 10 years, the p53 transcription factor that acts as a tumor suppressor has been identified as a key regulator of cell Sciences, Lyon, France 3Institute for Drug Research, the Hebrew bioenergetics metabolism. Many mutations of p53 result in the stabilization of mutant forms of the protein exerting University, Jerusalem, Israel a wide range of gain-of-function effects but little in known on how these effects impact on cell metabolism. Here, we investigate a collection of non-transformed fibroblasts cell lines (non-tumor cells) from different carriers of ger-mline TP53 mutations, associated with Li-Fraumeni Syndrome (LFS), a dramatic familial predisposition to cancer. We then detail the impact of short hairpin RNA (shRNA)-mediated p53 knockdown in these cells and associated control fibroblasts expressing wild-type p53, to gain specific insight into gain-of-function features associated with p53 mutations. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN051 PARALLEL SESSIONS / INVITED SPEAKER / METABOLOMICS 144 KAVITA DORAI USING NMR SPECTROSCOPY TO INVESTIGATE Indian Institute of Science THE LINK BETWEEN CIRCADIAN RHYTHMS AND Education & Research, Mohali, India METABOLISM The circadian clock controls several biological processes and is in turn influenced by environmental cues and by metabolic signals. Studies have shown that disruption in circadian rhythms can cause growth retardation in plants and metabolic disorders in fruit flies. We used NMR-based metabolomics to profile the metabolites in Drosophila melanogaster that cycle with a daily rhythm. The plant circadian clock is intimately connected with its response to abiotic stress and we used NMR methods for metabolite fingerprinting of the leaves of Bougainvillea spectabilis. We used NMR fingerprinting to understand how the sunflower plant’s circadian clock influences its solar tracking. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PT057 PARALLEL SESSIONS / PROMOTED TALK / METABOLOMICS 145 ŠTĚPÁN HORNÍK, NMR AEROSOLOMICS JAN SÝKORA, JAROSLAV SCHWARZ, Water-soluble organic compounds (WSOC) represent up to 80% identified. Three compounds were identified for the first time in VLADIMÍR ŽDÍMAL of all organic compounds present in atmospheric aerosols [1]. Un- ambient aerosol and the presence of other four species - predict- like composition of inorganic compounds or volatile organic com- ed by theory was confirmed. Altogether, the number of identified Institute of Chemical Process Fundamentals of the CAS, Prague, Czech pounds, which seems to be well explored, the knowledge about compounds climbed up to 79. The concentration levels found were Republic WSOC composition is still rather limited. The most frequently used subsequently used for statistical analysis. A clear discrimination method for WSOC analysis is GC-MS, which is a very sensitive tech-between summer and winter samples was achieved while the sep- nique [2]. However, the analysis of polar compounds via GC-MS aration according to particle size was indicated only in the group requires derivatization and the quantification is extremely time of summer samples. Univariate statistical analysis revealed sub-consuming. The second widely used technique is ion chromatog- stances responsible for the group separation and possible sources raphy (IC). Nowadays, IC is routinely used for analysis of specific of these compounds were suggested [5]. groups of organic compounds such as carboxylic acids, amines or Furthermore, the method was employed in WSOC analysis of size-re-carbohydrates [3]. NMR spectroscopy was for the purpose of aerosol solved aerosol particulate matter obtained by 6-stage high-volume chemistry employed only recently [4]. So far, the use of NMR spec- cascade impactor. Four series of samples collected in summer and troscopy is mainly restricted to so-called Functional Group Analy- winter were resolved into 6 fractions according to the particle size. sis, of which main interest lies elsewhere than in identification of In each sample, 31–45 compounds were identified, 73 compounds individual compounds. in total. The distribution profiles of individual compounds enabled NMR Aerosolomics offers a different approach to the analysis of identification of common sources or degradation pathways. INTRODUCTION complex aerosol mixtures inspired by compound profiling in me- Contrary to other methods, NMR aerosolomics allows determina- SPONSORS tabolomics. For this purpose, an extensive library of compounds tion of several groups of WSOC simultaneously. The method offers PROGRAM commonly present in aerosols was created. The method was eval- a great potential for source apportionment, which is a resonating PROGRAM — BY DAY uated on a series of 21 samples of PM2.5 and PM10 atmospheric topic in contemporary aerosol chemistry. TOPICS aerosol collected in Prague suburban site in summer 2008 and PRIZE LECTURES winter 2009. In each sample, approximately 60 compounds were PLENARY LECTURES TUTORIAL LECTURE References: [1] R. M. B. O. Duarte, A. Duarte, Trends Anal. Chem. 2011, 30, 1659-1671. [2] M. C. Pietrogrande, D. Bacco, Anal. Chim. Acta 2011, 689, 257-264. [3] Y. I. Tsai, L.Y. Hsieh, T. H. Weng, INVITED AND PROMOTED LECTURES Y. C. Ma, S. C. Kuo, Anal. Chim. Acta 2008, 626, 78-88. [4] S. Decesari, M. C. Facchini, S. Fuzzi, E. Tagliavini, J. Geophys. Res. 2000, 105, 1481–1489. [5] Š. Horník, J. Sýkora, J. Schwarz, V. Ždímal POSTERS ACS Omega 2020, 5, 36, 22750-22758. ADVERTISERS Acknowledgements: This work was supported by Technology Agency of the Czech Republic project (TK02010035) and by the infrastructure project of the Ministry of Youth, Education and AUTHOR INDEX Sports of the Czech Republic (LM2018122). PT058 PARALLEL SESSIONS / PROMOTED TALK / METABOLOMICS 146 EWA K. NAWROCKA1,2, VARIABLE-TEMPERATURE NMR SPECTROSCOPY - MATEUSZ URBAŃCZYK1, A HANDY TOOL FOR METABOLITE IDENTIFICATION IN KRZYSZOF KAZIMIERCZUK1 1Centre of New Technologies, University of BIOLOGICAL MATERIALS Warsaw, Warsaw, Poland 2Faculty of Chemistry, University of NMR spectroscopy is a beneficial method for identifying metabolites. Metabolic analysis is an essential part of the Warsaw, Warsaw, Poland modern diagnosis of many disease entities [1]. However, 1H-NMR spectra of metabolite mixtures (serum, plasma, urine ect.) usually used in medical screening are often very crowded and many peaks cannot be easily assigned to specific compounds. We can solve this problem by measuring 2D spectra but it is time-consuming [2], because of low concentrations and sampling requirements. In this work we exploit the fact, that chemical shift of metabolites change with temperature and that the rates of change (“temperature coefficients”) are characteristic for compounds and consistent between the samples. To determine them efficiently, we use variable-temperature (VT) NMR. When combined with Radon transform [3], VT-NMR allows measuring full VT series in a time comparable with a single 1H measurement. We show that certain metabolites (like alanine, lactate, L-valine, acetate, acetone, creatine, choline) have very specific and reproducible coefficients which can be used for their identification. We place hope in the fact, that VT-NMR can become an efficient tool for identifying chemical substances in biological materials and beyond. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Vignoli A. et al. , Angew. Chem. Int. Ed. Engl. 2019, 58(4), 968-994. [2] Kruk J., Doskocz M., Jodłowska E. et al. , Appl. Magn. Reson. 2017, 48, 1–21. POSTERS [3] Kupče E., Freeman R., J. Am. Chem. Soc. 2013, 135, 2871–2874. ADVERTISERS Acknowledgements: We thank the Foundation for Polish Science for support via the FIRST TEAM programco-financed by the European Union under the AUTHOR INDEX European Regional Development Fund no. (POIR.04.04.00-00-4343/17-00). PT059 PARALLEL SESSIONS / PROMOTED TALK / METABOLOMICS 147 L. T. KUHN,1 IN SITU ENANTIOSPECIFIC DETECTION OF K. MOTIRAM-CORRAL,2 MULTIPLE METABOLITES IN MIXTURES USING NMR T. J. ATHERSUCH,3 T. PARELLA,2 SPECTROSCOPY M. PÉREZ-TRUJILLO2 1 Chirality plays a fundamental role in nature, but its detection and quantification still face many limitations. To Institut für Physikalische Chemie, Albert-Ludwigs-Universität Freiburg, date, the enantiospecific analysis of mixtures necessarily requires prior separation of the individual components. Freiburg, Germany The simultaneous enantiospecific detection of multiple chiral molecules in a mixture represents a major challenge, 2Servei de Ressonància Magnètica which would lead to a significantly better understanding of the underlying biological processes; e.g. via enantio-Nuclear, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, specifically analyzing metabolites in their native environment. Here, we report on the first in situ enantiospecific Spain detection of a thirty-nine-component mixture. As a proof of concept, eighteen essential amino acids (AAs) at physi-3Department of Metabolism, Imperial College London, London, ological concentrations were simultaneously enantiospecifically detected using NMR spectroscopy and a chiral sol-UK vating agent. This work represents a first step towards the simultaneous multicomponent enantiospecific analysis of complex mixtures, a capability that will have substantial impact on metabolism studies, metabolic phenotyping, chemical reaction monitoring, and many other fields where complex mixtures containing chiral molecules require efficient characterization [1]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] L. T. Kuhn, K. Motiram-Corral, T. J. Athersuch, T. Parella, M. Pérez-Trujillo, Angew. Chem. Int. Ed. 2020, 59, 23615-23619. ADVERTISERS Acknowledgements: Financial support provided by the Spanish Ministry of Economy and Competitiveness (project PGC2018-095808-B-I00) is gratefully AUTHOR INDEX acknowledged. IN052 PARALLEL SESSIONS / INVITED SPEAKER / ORGANIC AND COMPOSITE SOLIDS 148 KLAUS SCHMIDT-ROHR, SPIN DIFFUSION AFTER INVERSION RECOVERY (SDAIR) ZHENHUAN SUN, FOR DETERMINATION OF 10 – 300 NM DOMAIN SIZES SHICHEN YUAN Department of Chemistry, Brandeis IN ORGANIC SOLIDS University, Waltham, MA, USA 1H spin diffusion after inversion recovery (SDAIR) is a simple solid-state NMR experiment that can provide accurate estimates of domain sizes in organic solids in the range of 10 – 300 nm, in favorable cases. SDAIR can be described as a one-dimensional 1H spin exchange experiment with 13C detection; selective excitation of protons in component A is achieved by inversion recovery to the zero-crossing of component B. 1H spin diffusion between the components during a mixing time of up to ~1500 ms results in distinctive exchange peaks of the initially nulled component B and an accelerated decrease of the retained signal A. While the intrinsic relaxation times need to differ by >30% for sufficient signal to be generated by SDAIR, the domain sizes can be obtained from peak intensities without knowledge of the intrinsic relaxation times, using versatile graphical analyses. The exchange peaks are largest for layer thicknesses of 40 – 80 nm, but can be detected for domains of up to 300 nm if the intrinsic relaxation times are sufficiently different. The experiment is demonstrated on PS- b-PMMA block copolymers. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX IN053 PARALLEL SESSIONS / INVITED SPEAKER / ORGANIC AND COMPOSITE SOLIDS 149 ANDRAŽ KRAJNC, SOLID-STATE NMR VIEW OF LOCAL STRUCTURE IN ZEOLITE BETA GREGOR MALI DESIGNED FOR FUTURE APPLICATIONS National Institute of Chemistry, Department of Inorganic Zeolite Beta is one of the most important zeolites in the industry. enthalpy. Additionally, the helical conformation of the perfluorinat-Chemistry and Technology, Ljubljana, Slovenia It is widely used for the reduction of VOC, N O and NO , industrial ed chains made a very tight packing at maximal capacity sterically 2 x gas purification, and automotive emission control. Its three-dimen- possible. This explained the exceptionally high affinity and selectiv- sional network of 12-ring pores makes it an ideal candidate for ad- ity of all-silica zeolite Beta for PFOA and similar perfluorinated com- sorption-based applications and also in catalysis for the production pounds, which positions this zeolite as the lead candidate for combat-of various chemicals, especially if shape-selectivity is desired. To ing the PFCs pollution. For utilization of zeolite Beta in catalysis, the understand the favorable properties of zeolites such as adsorption number of Brønsted acid sites and their strength play an important affinity, total uptake, selectivity, catalytic conversion, an insight into role. A higher Si/Al ratio represents fewer but stronger acid sites and the local structure of the framework is needed, and not many exper-makes pores more hydrophobic. We showed that a lower Si/Al ratio imental techniques are capable to offer it. In this regard, solid-state resulted in a higher conversion of citric acid to tricarballylic acid, evi-nuclear magnetic resonance is one of the most powerful techniques. dencing that in this reaction the number of acid sites is more import- ant than the strength of these sites. The Si/Al ratio before/after the We present zeolite Beta in its all-silica form, which shows outstand- reaction was determined from 29Si and 27Al MAS NMR spectra [2]. We ing capacity and affinity for the uptake of perfluorinated compounds noticed a decent increase in the ratio due to dealumination by citric (PFCs) such as perfluorooctanoic acid (PFOA) and perfluorooctane-acid. After recycling the zeolite through realumination, a significant sulfonic acid, even in the presence of other organic compounds. Re- fraction of the initial activity of the catalytic system was regenerated. lying mainly on 1H and 19F MAS NMR and 19F–29Si heteronuclear and INTRODUCTION On the contrary, a high Si/Al ratio was preferential for the shape-se- 1H–1H DQ-SQ homonuclear correlation experiments we elucidated SPONSORS lective conversion of simple aromatics to biarylic compounds using the way in which the PFOA molecules were adsorbed into the zeolite PROGRAM zeolite Beta loaded with 13C-enriched Pd(OAc) . 13C MAS NMR proved [1]. We showed that PFOA molecules were positioned in the straight 2 PROGRAM — BY DAY that one acetate out of the two react to form acetic acid, moving freely a- and b-channels of the zeolite, with the hydrophobic chains in the TOPICS in the pores, while the second acetate remains immobilized at a Pd channels and the carboxylic heads, forming hydrogen-bonded pairs, PRIZE LECTURES centre which is strongly bound to the framework aluminum [3]. in the intersections. This resulted in a very favorable adsorption PLENARY LECTURES TUTORIAL LECTURE References: [1] M. Van den Bergh, A. Krajnc, S. Voorspoels, S. R. Tavares, S. Mullens, I. Beurroies, G. Maurin, G. Mali, D. E. De Vos, Angew. Chem. Int. Ed. 2020, 59, 14086–14090. [2] W. Stuyck, INVITED AND PROMOTED LECTURES J. Verduyckt, A. Krajnc, G. Mali, D. E. De Vos, Green Chem. 2020, 22, 7812–7822. [3] J. Vercammen, M. Bocus, S. Neale, A. Bugaev, P. Tomkins, J. Hajek, S. Van Minnebruggen, A. Soldatov, A. POSTERS Krajnc, G. Mali, V. Van Speybroeck, D. E. De Vos, Nat. Catal. 2020, 3, 1002–1009. ADVERTISERS Acknowledgements: We thank FWO (research grant G0781118N, G0D0518N) and Slovenian Research Agency for the financial support (research core funding P1-0021 and research projects AUTHOR INDEX N1-0079 & Z1-9171). PT060 PARALLEL SESSIONS / PROMOTED TALK / ORGANIC AND COMPOSITE SOLIDS 150 MARTIN DRAČÍNSKÝ, MOLECULAR MOTORS STUDIED BY SOLID-STATE NMR CARINA SANTOS HURTADO, SPECTROSCOPY GUILLAUME BASTIEN, JIŘÍ KALETA Artificial molecular machines promise applications in many fields, including physics, information technologies, Institute of Organic Chemistry and chemistry as well as medicine. The deposition of functional molecules in 2D or 3D assemblies in order to control Biochemistry, Czech Academy of Sciences, their collective behavior and the structural characterization of these assemblies are challenging tasks. We exploit Prague, Czech Republic; dracinsky@uochb.cas.cz porous materials to form rigid matrix for mechanochemical preparation of bulk or surface host−guest inclusions with functional molecules, such as molecular rotors, molecular motors and molecular switches. Unambiguous determination of the molecular structure and monitoring of the molecular function such as rota- tion of a molecular rotor or on/off switching of a molecular switch cannot be studied by X-ray analysis because the systems are typically heavily disordered fine powders. We use solid-state nuclear magnetic resonance (SS-NMR) spectroscopy to obtain atomic-level insights into the structure and dynamics of these functional materials. SS-NMR spectra provide valuable information about structure, interactions and dynamics in solids not available otherwise. It will be demonstrated that SS-NMR experiments provide unequivocal evidence of the formation of the 2D and 3D assemblies and can also be used for the observation of such a molecular function as the photoisomerization of a molecular switch deposited on a surface. We have also developed a solid-state NMR method for investigation of two dimensional arrays of light-driven molecular motors [1-4]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] J. Kaleta, J. Chen, G. Bastien, M. Dračínský, M. Mašát, C. T. Rogers, B. L. Feringa, J. Michl, J. Am. Chem. Soc. 2017, 139, 10486. [2] J. Kaleta, G. INVITED AND PROMOTED LECTURES Bastien, J. Wen, M. Dračínský, E. Tortorici, I. Císařová, P. D. Beale, C. T. Rogers, J. Michl, J. Org. Chem. 2019, 84, 8449. [3] C. Santos-Hurtado, G. Bastien, M. Mašát, J. R. POSTERS Štoček, M. Dračínský, I. Rončević, I. Císařová, C. Rogers, J. Kaleta, J. Am. Chem. Soc. 2020, 142, 9337. [4] M. Dračínský, C. Santos-Hurtado, E. Masson, J. Kaleta, ADVERTISERS Chem. Commun. 2021, 57, 2132. AUTHOR INDEX Acknowledgements: This work was supported by the Czech Science Foundation (grant no. 20-01472S). PT061 PARALLEL SESSIONS / PROMOTED TALK / ORGANIC AND COMPOSITE SOLIDS 151 MANUEL CORDOVA1,2, BAYESIAN PROBABILISTIC ASSIGNMENT MARTINS BALODIS1, OF ORGANIC SOLIDS BRUNO SIMÕES DE ALMEIDA1, The starting point for any detailed NMR study is the assignment of the experimental NMR spectra. In organic solids MICHELE CERIOTTI2,3, at natural isotopic abundance, this is still a laborious and often challenging process. In solution, when required, LYNDON EMSLEY1,2 probabilistic assignments can be obtained without prior knowledge of the three-dimensional structure through a 1Laboratory of Magnetic Resonance, statistical analysis of large experimental chemical shift databases [1]. However, no such database exists for organic Institute of Chemical Sciences and solids. Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland 2 Here, we obtain the chemical shifts of over 200 000 organic crystal structures obtained from the Cambridge struc-National Centre for Computational Design and Discovery of Novel Materials MARVEL, tural database [2] using ShiftML, a machine learning model able to predict chemical shifts in molecular solids [3], in Ecole Polytechnique Fédérale de Lausanne order to construct a statistical basis for the probabilistic assignment of solid-state NMR spectra. Relating the topo- (EPFL), Lausanne, Switzerland 3Laboratory of Computational Science and logical representation of molecular fragments to their corresponding statistical distribution of predicted chemical Modelling, Institute of Materials, Ecole shifts, we propose a Bayesian probabilistic framework to determine possible assignments of experimental NMR Polytechnique Fédérale de Lausanne (EPFL), spectra of organic crystals, along with their confidence, from the two-dimensional representation of these com- Lausanne, Switzerland pounds only. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] T. Aeschbacher, E. Schmidt, M. Blatter, C. Maris, O. Duss, F. H. T. Allain, P. Güntert, M. Schubert, Nucleic acids res 2013, 41, e172. [2] C. R. Groom, POSTERS I. J. Bruno, M. P. Lightfoot, S. C. Ward, Acta Crystallogr. B 2016, 72, 171. [3] F. M. Paruzzo, A. Hofstetter, F. Musil, S. De, M. Ceriotti, L. Emsley, Nat. Commun. ADVERTISERS 2018, 9, 4501 AUTHOR INDEX Acknowledgements: SNSF grant no. 200020_178860 and the NCCR MARVEL. PT062 PARALLEL SESSIONS / PROMOTED TALK / ORGANIC AND COMPOSITE SOLIDS 152 BING WU1,2,3, SOLID STATE NMR PROBING POLYMER CHAIN MOBILITY KISHAN ARYASOMAYAJULA2, AT DIFFUSED INTERPHASES FORMED IN FARHAD SHAHSAVAN4, HAIYAN MAO2, MULTI-MICRO-/NANOLAYERED PVDF-PMMA FILMS JEFFREY A. REIMER2, ALEXEY KRUSHELNITSKY4, For multi-layered polymers, polymer–polymer interfaces play a Solid-state nuclear magnetic resonance (NMR) is one of the most DANIEL HERMIDA MERINO3, crucial role in the final properties of these multiphase materials. powerful techniques for elucidating details of segmental dynamics KAY SAALWÄCHTER4, Among all the techniques for producing multi-layered films, the in solid materials. While fast dynamics can be characterized by tech-ANDREAS HEISE1 coextrusion process is one of the most appealing and popular tech- niques like line shape analyses or relaxation measurements [1], some 1 niques for industrial scale processing. This process is widely used specific dynamic information can only be obtained in exchange NMR Royal College of Surgeons in Ireland, Department of Chemistry, Dublin, Ireland to form multi-layered sheets or films that are suitable for various experiments, where relatively slow segmental reorientations are ob-2University of Californi Berkeley, products ranging from food packaging materials to dielectric capac- served in terms of changes of orientation-dependent NMR frequen- Department of Chemical and Biomolecular itors. Using layer multipliers or layer-multiplying elements, prod- cies. The centerband-only detection of exchange (CODEX) NMR tech- Engineering, Berkeley, California, USA 3European Synchrotron Radiatin Facility, ucts with thousands of layers can be produced, in which the layer nique introduce by Schmidt-Rohr group is one of these exchange Dutch Belgium Beamline, Grenoble, France thickness can be reduced to the nanometre scale (Figure 1a). Inter- NMR technique, which is possible to observe and characterize very 4Martin Luther University Halle-Wittenberg, Instit für Physik, Halle, Germany facial spatial confinement always dominates when layer thickness slow segmental reorientations with the highest available NMR senis decreased, which greatly alters the microstructure and dynamics sitivity and site resolution, in sideband free magic-angle spinning of the multilayer polymers. Macroscopic proper ties, including me- (MAS) spectra. From short series of one-dimensional MAS spectra, chanical, electric, and gas/liquid barrier properties, are also drasti- the correlation function and correlation time can be determined. In cally altered. In addition, for the nanolayer coextrusion process, the this study, a series of multi-micro/nanolayered PVDF/PMMA films INTRODUCTION laminar flow conditions combine polymers in the layer multipliers were analyzed by CODEX at 4 different temperatures. It is found that SPONSORS by producing a large number of interlayer interfaces without com- The a -relaxation of PVDF accelerated upon decreasing the layer a PROGRAM pleted mixing. Interfacial behaviours involving interlayer diffusion thickness, while the a relaxation does not shift significantly. These c PROGRAM — BY DAY and reactions are also critical in defining the structure and proper- results are further discussed in relation to DRS analyses and MD sim- TOPICS ties. The measurement of the interfacial properties is important for ulation [2]. The effects of the amount and orientation of crystalline PRIZE LECTURES understanding the interdependence of processing, structure, and phase on this suppression are also discussed via SAXS/WAXS and PLENARY LECTURES properties. DSC analyses of these samples. TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Roos, M. et al., Colloid. Polym. Sci. 2014, 292 (8), 1825-1839. [2] Shi, M. N. et al., J. Phys. Chem. B 2016, 120 (37), 10018-10029. AUTHOR INDEX Acknowledgements: Financial support from European Commission Marie Skłodowska-Curie Actions. 153 POSTERS PO001 POSTERS / BIOMOLECULES AND INTERACTIONS 154 ADLER AGNES1, AN NMR VIEW OF DYNAMIC MICROTUBULE-MAP LUO YANZHANG1,2, INTERACTIONS BALDUS MARC1 1NMR Spectroscopy group, Microtubules (MT) play an essential role in cell migration, mitosis and polarization. Many of these functions criti-Bijvoet Center for Biomolecular cally rely on Microtubule-associated proteins (MAPs) and their association with microtubules. So far, little is known Research, Utrecht University, Utrecht, The Netherlands about the interaction of MAPs and their intrinsically disordered regions with the dynamic microtubule surface. 2MAGNEtic resonance research In particular, the role of the unstructured C-terminal tails of tubulin, critical for many MAPs interactions, has re-FacilitY (MAGNEFY), Wageningen mained elusive. Considering the importance of protein dynamics for MT function, and especially the role of the University & Research, Stippeneng, The Netherlands highly flexible tubulin tails in cellular processes and human disorders such as ciliopathies, cancer and neurodegeneration [1], we have employed nuclear magnetic resonance (NMR) to obtain structural and dynamical information about these interactions at atomic level [2,3,7]. Using a combination of solution- and solid-state NMR spectroscopy, we have examined the interaction of MT with different MAPs. In our studies, we made use of MT protofilaments, Tubulin-dimers and peptides including those representing the tubulin C-terminal tails. Specific experiments were designed to help probe interaction between the MAPs and the tubulin C-terminal tails. In our studies, we focused on three different members of the MAP family. [1] The CKK domain of the calmodulin-regulated spectrin-associated protein (CAMSAP), involved in the minus-end recognition of the MT [3,4]. MAP7 which is essential in regulating kinesin-based intracellular transport and competes with the MAP tau for MT-binding [3,6]. Lastly, we examined the binding mechanism of the Alzheimer’s dis- ease-related MAP Tau to MT [5]. INTRODUCTION SPONSORS In our contribution, we report on progress on combining our NMR results with information obtained by biochemi- PROGRAM cal and biophysical methods to gain deeper insight into the binding behavior of these MAPs to MT. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Gadadhar, S., Bodakuntla, S., Natarajan, K. & Janke, C. J. Cell Sci. 2017, 130, 1347–1353. [2] Atherton, J. et al. , Nature Communications, 2019, 10, 1-16. [3] Luo, Y. et al. , Nature Communications 2020, 11, 18. [4] Atherton, J. et al. Nature Structural & Molecular Biology 2017, 24, 931. [5] Medeiros R, Baglietto- INVITED AND PROMOTED LECTURES Vargas D, LaFerla FM. CNS neuroscience & therapeutics 2011, 17, 514-24. [6] Monroy, B. Y. et al. Nat. Commun. 2018, 9, 1487. [7] Luo Y, Xiang S, Paioni AL, et al. POSTERS Methods Mol Biol. 2021, 2305, 193-201. ADVERTISERS Acknowledgment: We thank Joseph Atherton, Peter Jan Hooikaas, Anna Akhmanova and Carolyn Moores for giving further insights into this research topic AUTHOR INDEX by Cryo-EM and molecular biology. PO002 POSTERS / BIOMOLECULES AND INTERACTIONS 155 SIMON ALEKSIČ1, 8-OXOGUANINES ARE ACCOMMODATED IN JANEZ PLAVEC1,2,3, A G-QUADRUPLEX QUARTET PETER PODBEVŠEK1 1National Institute of Chemistry, Reactive oxygen species (ROS) are a by-product of cellular metab- model sequence. Using NMR spectroscopy, we determined the abil- Slovenian NMR Centre, olism in all living organisms. Cells posses (non)-enzymatic anti- ity of the synthetized oligonucleotides to fold into G-quadruplexes Ljubljana, Slovenia 2EN-FIST Centre of Excellence, oxidant mechanisms, which are able to maintain normal levels of and proved all four investigated oligonucleotides formed G-quadru-Ljubljana, Slovenia endogenous ROS. Since guanine has the lowest redox potential of plexes. We investigated the thermal stability of the G-quadruplexes 3University of Ljubljana, Faculty the four nucleobases, it is highly susceptible to oxidation damage. with NMR melting experiments. Using two-dimensional through-of Chemistry and Chemical Technology, Ljubljana, Slovenia One such oxidation product is 8-oxoguanine [1]. Guanine rich re- bond and through-space NMR methods, we have confirmed the par- gions are present in the promoter and telomere regions of genomes allel topology of all formed G-quadruplexes. The accommodation of and are able to fold into noncanonical four-stranded structures 8-oxoguanine moieties into a quartet was explored using simulat-called G-quadruplexes It is presumed that G-quadruplex forming ed annealling molecular modelling. We confirmed a possibility of sequences act as oxidation sinks in the genome. 8-Oxoguanine for-hydrogen bond formation between the amino group hydrogen and mation can lead to nucleic acid structural rearrangements and can oxygen on position 8 between neighbouring 8-oxoguanine residues have an effect on cellular mechanisms [2,3]. Insights into the effect in a quartet. of 8-oxoguanine incorporation on G-quadruplex structure will help With the attained data, we conclude that four 8-oxoguanine moi- expand the knowledge of ROS damage on nucleic acids and the eties are accommodated in an all-8-oxoguanine quartet, noticeably structural changes that are a consequence of oxidative stress. impacting the thermal stability of G-quadruplexes only when po- We have chosen a short model deoxyoligonucleotide TG T that is sitioned inside the quadruplex core. The knowledge attained will 4 INTRODUCTION able to fold into a tetrameric G-quadruplex in solution. We have guide further studies on effect of 8-oxoguanine formation on long SPONSORS residue-specifically incorporated 8-oxo-deoxyguanosine into the guanine-rich sequences. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] A. M. Fleming, C. J. Burrows, DNA Repair, 2017, 56, 75-83. [2] M. Vorlícková, M. Tomasko, A. J. Sagi, K. Bednarova, J. Sagi, FEBS J. , 2012, 279, 29-39. [3] S. C. J. Redstone, A. M. ADVERTISERS Fleming, C. J. Burrows, Chem Res. Toxicol. , 2019, 32, 437-446. AUTHOR INDEX Acknowledgements: This work is part of the PhD “The effect of oxidative stress on guanine rich genomic regions” supported by CERIC-ERIC and ARRS research programme P1-0242. PO003 POSTERS / BIOMOLECULES AND INTERACTIONS 156 CARLA GARCIA-CABAU1,2, CHARACTERIZATION OF THE LIQUID-LIQUID PHASE LORENZO BRACAGLIA3, SEPARATION OF THE PROGESTERONE RECEPTOR ELZBIETA SZULC1,2, JESÚS GARCÍA1,2, ACTIVATION DOMAIN USING ITS SEQUENCE-SPECIFIC ROBERTA PIERATTELLI3, RESONANCE ASSIGNMENT ISABELLA C. FELLI3, XAVIER SALVATELLA1,2,4 The uman progesterone receptor (PR) is a ligand-activated transcription factor member of the steroid receptor su-1Institute for Research in Biomedicine (IRB perfamily. The isoform B counts 933 residues and it is composed of a proline-rich intrinsically disordered (ID) activa-Barcelona), The Barcelona Institute of tion domain (AD), followed by globular DNA- and ligand-binding domains. PR AD consists of 566 residues, and it is Science and Technology (BIST), Barcelona, Spain reported to play a crucial role for protein function [1]. It has recently been shown that transcription can be regulated 2Joint BSC-IRB Research Programme in by the liquid-liquid phase separation (LLPS) of the transcription machinery, stabilized by interactions between the Computational Biology, Barcelona, Spain 3 ID regions of transcription factors [2]. Magnetic Resonance Center and Department of Chemistry “Ugo Schiff”, University of Florence, Sesto Fiorentino In this work we have characterized the LLPS properties of PR AD by correlating the compaction degree of the mono- (Florence), Italy mer with the propensity to undergo LLPS. In order to get residue level information to identify the regions of se-4ICREA, Barcelona, Spain quence driving this process, we used NMR and carried out the sequence-specific resonance assignment of the dis- ordered domain through the application of a mixed set of 1H-detected and 13C-detected NMR experiments [3]. With this approach, the N-terminal domain has been almost completely assigned, including the proline residues that represent the 15% of the sequence. With this information, we then assessed the chemical shift perturbations and intensity changes that different parameters that influence LLPS have on the PR AD spectra. INTRODUCTION SPONSORS In conclusion, we have found out that PR AD undergoes LLPS in vitro and that the formed liquid droplets are stabi- PROGRAM lized by a mixture of electrostatic and hydrophobic interactions. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] G. S. Takimoto et al. , J. Steroid Biochem. Mol. Biol. 2003, 85, 209–219; [2] Hnisz, D., Abraham, B.J., Lee, T.I., Lau, A., Saint-André, V., Sigova, A.A., AUTHOR INDEX Hoke, H.A., Young, R.A. Cell 2013, 155, 934–947; [3] I.C. Felli and R. Pierattelli, J. Magn. Reson. 2014, 241,115-25. PO004 POSTERS / BIOMOLECULES AND INTERACTIONS 157 MIRKO CEVEC1, ANALYSIS OF NRAS RNA G-QUADRUPLEXES JANEZ PLAVEC1,2,3 1 RNA guanine-rich sequences are able to form G-quartets through hydrogen bond interactions in the presence of National Institute of Chemistry, Slovenian NMR Centre, monovalent cations such as potassium and sodium ion. G-quartets stack onto each other and assemble G-quadru- Ljubljana, Slovenia plexes, which have broad diversity regarding loop lengths and arrangements, but they commonly adopt the parallel 2EN-FIST Centre of Excellence, conformation in which all four strands are aligned in the same direction. RNA G-quadruplexes are dynamic, tran- Ljubljana, Slovenia 3University of Ljubljana, Faculty sient in cells, and are included in diverse biological processes, such as translation, regulation of alternative splicing, of Chemistry and Chemical and the subcellular transport of mRNAs. They are used as therapeutic targets and agents. Technology, Ljubljana, Slovenia RNA G-quadruplex forming sequence in the 5’ untranslated region (UTR) of neuroblastoma RAS viral oncogene homo-logue ( NRAS) proto-oncogene was studied here. NRAS proto-oncogene encodes for a protein called N-Ras, which is involved in regulating cell division through signal transduction. We will present our results on RNA G-quadruplex structures in the 5’-UTR of the NRAS mRNA utilizing NMR, CD, and UV spectroscopies. We also synthesized modified RNA G-quadruplexes with different G-tract and loop lengths in order to stabilize the most possible structures. Our data show that modified sequences form one structure. NRAS RNA G-quadruplexes have parallel connectivity with propeller loops. Structure with G5 in the loop is the most probable. Results will help to develop new tools to use NRAS RNA G-quadruplexes as therapeutic targets. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PO005 POSTERS / BIOMOLECULES AND INTERACTIONS 158 NASTJA FEGUŠ1,2, CAN MATURE LET-7 MICRORNAS FORM JANEZ PLAVEC1,2,3, NONCANONICAL SECONDARY STRUCTURES? ANITA KOTAR1 1National Institute of Chemistry, The human genome contains thousands of non-coding (nc)RNAs that are transcribed from DNA but are not trans- Slovenian NMR Centre, lated into proteins [1]. It has been shown that ncRNAs are involved in many normal cellular processes as well as Ljubljana, Slovenia 2University of Ljubljana, Faculty disease states [1]. Herein, we focused on microRNAs (miRNAs), a group of ncRNAs, which play important roles in of Chemistry and Chemical the regulation of gene expression through targeting messenger (m)RNAs for post-transcriptional gene silencing Technology, Ljubljana, Slovenia 3 [2]. Specifically, we are studying letal-7 miRNA (let-7) family which has two major biological functions in cells, 1) as EN-FIST Centre of Excellence, Ljubljana, Slovenia essential regulators of differentiation and 2) as fundamental tumor suppressors [3]. In humans, the let-7 family is composed of nine isoforms of mature RNAs (let-7a, let-7b, let-7c, let-7d, let-7e, let-7f, let-7g, let-7i, and miRNA-98). It was shown before that stable secondary structures, such as G-quadruplexes, in mature miRNAs can affect base pairing with mRNA and thus alter gene regulation [4]. Our NMR data revealed that all studied members of the let-7 family, except let-7a and miRNA-98, form stable secondary structures. Interestingly, three of them, let-7b, let-7c, and let-7e, adopt G-quadruplexes as indicated by characteristic imino signals in the region between 10.5 and 11.9 ppm observed in 1D 1H NMR spectra. This observation was unexpected because although these RNAs are guanine (G) rich, they do not conform to a classical G-quadruplex motif composed of four runs of three(two) Gs. The secondary structures of the mature let-7 family were examined by CD spectroscopy which revealed that let-7 G-quadruplex structures adopt parallel topologies. The band migration on native polyacrylamide gels suggested the potential formation of multimeric G-quadruplexes, which was further explored by UV melting experiments. Interestingly, our INTRODUCTION preliminary NMR data demonstrate how mature let-7 miRNAs structures influence the base pairing with target SPONSORS mRNA. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] S. Djebali, C. A. Davis, A. Merkel, A. Dobin, T. Lassmann, et al. Nature. 2012 489, 101-108. [2] A. Zampetaki, A. Albrecht, K. Steinhofel, Front. POSTERS Physiol. 2018, 9:1201. [3] H. Lee, S. Han, C. S. Kwon, D. Lee, Protein Cell. 2016, 7, 100-113. [4] M. Tassinari, S. N. Richter, P. Gandellini., Nucleic Acids Res. 2021, ADVERTISERS 9, 3617–3633. AUTHOR INDEX Acknowledgments: Authors acknowledge the financial support of the Slovenian Research Agency [ARRS, grant P1-0242]. PO006 POSTERS / BIOMOLECULES AND INTERACTIONS 159 MARTINA LENARČIČ ŽIVKOVIĆ1,2, FORMATION PROPENSITY OF PSEUDOCIRCULAR DNA MARTIN GAJARSKÝ1,3, KATEŘINA G-HAIRPINS BEKOVÁ1,3, PETR STADLBAUER4, LUKÁŠ VICHEREK1, MAGDALENA PETROVÁ5, As we have recently shown, Saccharomyces cerevisiae telomeric DNA can fold into an unprecedented pseudocircular RADOVAN FIALA1, IVAN ROSENBERG5, G-hairpin (PGH) structure [1]. However, PGH formation in the context of extended sequences (a prerequisite for JIŘÍ ŠPONER1,4, JANEZ PLAVEC2,6,7, their function in vivo and their applications in biotechnology) has not been elucidated. LUKÁŠ TRANTÍREK1 1 Herein, we show that the structure‘s circular nature tolerates single-stranded (ss) protrusions. The high-resolution Central European Institute of Technology, Masaryk University, Brno, Czech Republic NMR structure of prolonged sequence revealed a new member of the PGH family and showed atomistic details 2National Institute of Chemistry, Slovenian NMR Centre, on a junction between ssDNA and structured PGH part. We also identified new sequences capable of folding into Ljubljana, Slovenia 3 one of the two forms of PGH and defined minimal sequence requirements for their formation. The time-resolved National Centre for Biomolecular Research, Masaryk University, Brno, Czech Republic 4Institute of Biophysics of NMR data revealed a possibility that PGHs fold via a complex kinetic partitioning mechanism. These data not only the Czech Academy of Sciences, Czech Republic explain its cation-type-dependent formation but also explain the unusual hysteresis between PGH melting and an- 5Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic nealing described in our previous study. Our findings have important implications for DNA biology and nanotech- 6EN-FIST Centre of Excellence, Ljubljana, Slovenia nology. Overrepresentation of sequences able to form PGHs in the evolutionary-conserved regions of the human 7Faculty of Chemistry and Chemical Technology, University genome implies their potential functionally important biological role(s) [2]. of Ljubljana, Slovenia INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Gajarský, M. et al., J. Am. Chem. Soc. 2017, 139, 3591–3594. [2] Živković, M. L. et al., Nucleic Acids Res. 2021, 49, 2317–2332. INVITED AND PROMOTED LECTURES Acknowledgments: Czech Science Foundation [19-26041X to L.T., 17-12703S to I.R.]; ‘MSCAfellow2@MUNI’ [CZ.02.2.69/0.0/0.0/18.070/0009846 to M.L.Z.]; POSTERS SYMBIT [CZ.02.1.01/0.0/0.0/15 003/0000477 to J.S.] financed by the European Regional Development Fund with financial contribution from the Ministry ADVERTISERS of Education, Youth, and Sports of the Czech Republic (MEYS CR); Slovenian Research Agency [P1-242, J1-1704 to J.P.]; CERIC-ERIC Consortium and projects AUTHOR INDEX CEITEC 2020 [LQ1601]; National Programme for Sustainability II; CIISB [LM2018127] (co-)funded by MEYS CR for access to experimental facilities. PO007 POSTERS / BIOMOLECULES AND INTERACTIONS 160 AGRIM GUPTA1, NMR DIRECTED MUTATIONS MODULATE CHRISTIAN MANUEL KITZLER1, COILED-COIL INTERACTIONS OF CALCIUM CHANNEL HERWIG GRABMAYR2, ADRIANA RATHNER3, PROTEINS PETR RATHNER1,4, MARC FAHRNER2, We investigate STIM1 the calcium sensor protein involved in the activation of the calcium released activated calci-MATTHIAS BECHMANN1, um (CRAC) channel. STIM1 spans from the endoplasmic reticulum into the cytosol [1]. It undergoes homo-oligo- CHRISTOPH ROMANIN2, AND merization and spatial elongation once the Ca2+ store is depleted, resulting in what is known as “store operated” NORBERT MÜLLER1,4 activation of the Orai Ca2+ channel. One long (CC1) and two short (CC2, CC3) coiled-coil segments are found in the 1 cytosolic part of STIM1, and they are involved in various intra- and intermolecular interactions transitioning be-Institute of Organic Chemistry, Johannes Kepler University Linz, Austria tween the resting and active states [2]. 2Institute of Biophysics, Johannes Kepler University Linz, Austria In its monomeric form, the isolated STIM1-CC1 domain in isotropic solution was shown to form a three-helix bun- 3Institute of Inorganic Chemistry, Johannes dle and stabilized mainly by two pairs of interhelical coiled-coil epitopes. In the Stormorken disease-related STIM1 Kepler University Linz, Austria 4 R304W mutant these contacts are weakened, according to our new NMR-based solution structure model. Two in- University of Vienna, Institute of Analytical chemistry, Vienna, Austria terhelical sites inside the domain between CC1a1 and CC1a2 subdomain helices have been identified as critical for 5Faculty of Science, University of South modulating channel activation [3]. Point mutations within interhelical interaction contact locations derived from Bohemia, České Budějovice, Czech Republic NMR structure and dynamics data can restore the physiological, store-dependent activation behavior of injurious mutant in living cells. The NMR-directed design of mutations within coiled-coil domains where interhelical interactions a1-a2 of CC1 compete with CC1-CC3 molecular “clamp” to control the function of STIM1 and recover func- INTRODUCTION tionality lost through other pathogenic mutations is demonstrated. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] J. Liou, M.L. Kim, W.D.Heo, J.T. Jones, J.W. Myers, JE jr. Ferrell, T. Meyer, Curr Biol 2005, 15, 1235-1241. [2] M. Muik, M. Fahrner, R. Schindl, TUTORIAL LECTURE P. Stathopulos, I. Frischauf, I. Derler, P. Plenk, B. Lackner, K. Groschner, M. Ikura, C. Romanin, The EMBO journal 2011, 30(9), 1678-89. [3] P. Rathner, M. Fahrner, L.Cerofolini, H. Grabmayr, F. Horvath, H. Krobath, A. Gupta, E. Ravera, M. Fragai, M. Bechmann, T. Renger, C. Luchinat, C. Romanin, N. Müller, Nature INVITED AND PROMOTED LECTURES Chemical Biology 2020, 17, 196-204. POSTERS Acknowledgments: The authors are grateful for financial support through the DK- NanoCell project by the Austrian Science Fund (FWF) W1250. All NMR ADVERTISERS experiments were performed at the NMR laboratory of the Upper Austrian-South Bohemian Research Infrastructure Center in Linz, co-financed by the AUTHOR INDEX European Union in the context of the project “RERI- uasb”, EFRE RU2-EU-124/100-2010 (ETC Austria-Czech Republic 2007–2013, project M00146). PO008 POSTERS / BIOMOLECULES AND INTERACTIONS 161 SIGURD RAMANS HARBOROUGH1, ORDER IN DISORDER: AUX/IAA PROTEIN AND ITS TIR1- ARNOUT P. KALVERDA1, GARY S. AUX/IAA AUXIN CO-RECEPTOR SYSTEM THOMPSON2, MARTIN KIEFFER1, MARTIN KUBES3, MUSSA QUARESHY3, Auxin is a central signaling molecule in plant biology with roles in both the patterning of developmental events and VESELINA UZUNOVA3, JUSTYNA M. the regulation of cellular growth. This is achieved via the TIR1/AFB-auxin-Aux/IAA co-receptor complex. Within PRUSINSKA3, KEN-ICHIRO HAYASHI4, this ternary complex, auxin acts as a molecular glue to promote binding of Aux/IAA transcriptional repressor pro-RICHARD NAPIER3, IAIN W. MANFIELD1, teins to SCFTIR1/AFB ubiquitin-ligase complexes, thereby catalyzing their ubiquitin-mediated proteolysis. A conspicu-STEFAN KEPINSKI1 ous feature of the crystal structure of the complex is a rare cis W-P bond within the binding site. This binding site is 1Centre for Plant Sciences and Astbury Centre for Structural centered on a 13 amino acid motif called the degron. We have used NMR to determine the solution structure of the Molecular Biology, Faculty of Biological Sciences, University amino-terminal half of the Aux/IAA-protein AXR3/IAA17 and its binding in complex with TIR1 and auxin. We show of Leeds, Leeds, United Kingdom 2School of Biosciences, University of Kent, Canterbury, United that this is intrinsically disordered and yet the critical degron W-P bond occurs with an unusually high (1:1) ratio of Kingdom cis to trans isomers. While the WPP sequence is one of the most strongly cis proline promoting elements. In peptides 3School of Life Sciences, University of Warwick, Coventry, containing this motif, the population of the cis conformer is at most 36%. This demonstrates that there must be at United Kingdom 4Departemnt of Biochemistry, Okayama University of least a transient structural element promoting the population of the cis conformer beyond this. Analysis of RDC’s Science, Okoyama, Japan confirms a deviation of random coil structure both in the degron motif and near the N-terminus, where a transient helix is formed that provides a key interface for the recruitment of the co-repressor TOPLESS. We then show that assembly of the co-receptor complex involves both auxin-dependent and -independent interaction events. Further, using the synthetic auxin molecule cvxIAA we show that a subset of auxin-dependent binding events occur away from the base of the canonical auxin binding pocket in TIR1. Our results suggest the existence of an encounter com- INTRODUCTION plex before the formation of the fully docked ternary complex and the involvement of the region C-terminal to the SPONSORS degron in auxin independent interactions with TIR1. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS Acknowledgements: We acknowledge the Astbury BioStructure Laboratory (ABSL) for access to the 950 MHz spectrometer which was funded by the ADVERTISERS University of Leeds. The ABSL NMR, Cryo-EM and MassSpec facilities are a member of INSTRUCT-ERIC and accept applications for measurement time from AUTHOR INDEX European based scientists. Contact the presenter for more information. PO009 POSTERS / BIOMOLECULES AND INTERACTIONS 162 PIOTR KLUKOWSKI, LEVERAGING DEEP LEARNING FOR AUTOMATED PROTEIN ROLAND RIEK, STRUCTURE DETERMINATION WITH NMR SPECTROSCOPY PETER GÜNTERT ETH Zurich, Department Nuclear magnetic resonance spectroscopy (NMR) is one of the lead- The above dataset has been used to train a deep residual neural of Chemistry and Applied ing techniques for protein structure determination. Nonetheless, network with 26 layers and over 3.5 million parameters, which per-Biosciences, Zurich, Switzerland the full potential of NMR spectroscopy remains unfulfilled due forms automated peak picking and peak deconvolution. Signals to the tedious data analysis process. Nowadays, it takes weeks or identified by the model are passed to automated assignment with months of manual work to deliver a structural model of the protein FLYA [1], yielding an initial chemical shift list. Subsequently, we out of measured NMR spectra. use another module of our workflow, namely a deep graph neural network (GNN). This model has been trained on shift lists extract- In this project, we have addressed the problem of fully automated ed from 2840 BMRB records. Its role is to capture short- and long-protein structure determination with deep learning. Our method range patterns that are present in protein chemical shifts. The GNN takes as input only the protein sequence and NMR spectra, produc- model brings domain knowledge from previously solved proteins to ing as output: (a) peak lists for each spectrum, (b) a chemical shift our workflow, allowing to refine the outputs of FLYA and automated list, (c) upper distance limit restraints, and (d) a protein structure in peak picking. The final step in our approach is automated NOESY PDB format. The structure determination process does not require assignment and structure calculation with CYANA [2]. any human intervention and takes about 5 hours, making it possi- ble to obtain a high-quality protein structure shortly after complet- Using our approach, we have managed to automatically solve 100 ing the NMR measurements. protein structures with a median backbone RMSD of 1.27 Å to the PDB reference structures. Moreover, the method correctly assigned INTRODUCTION A key factor for the successful delivery of deep learning solutions 96.3% backbone and 85.5% side-chain chemical shifts (median ac- SPONSORS is the availability of large-scale training datasets. We have collect- curacy), compared to BMRB depositions. Our method will be pub- PROGRAM ed 1 314 NMR spectra (2D, 3D, and 4D experiments), allowing to re- licly available this year as a website, which allows for spectra upload PROGRAM — BY DAY produce 100 protein structures of 35–175 residues, which have been and rapid protein structure determination. TOPICS solved manually in the past. In addition, we have prepared a bench- PRIZE LECTURES mark dataset, composed of 3 925 370 peak examples, including PLENARY LECTURES 650 675 manually labeled signals (true peak vs. artifact) and 13 831 TUTORIAL LECTURE signal deconvolution examples. INVITED AND PROMOTED LECTURES POSTERS References: [1] E. Schmidt, P. Güntert, J. Am. Chem. Soc. 2012, 134, 12817–12829. [2] P. Güntert, L. Buchner, J. Biomol. NMR 2015, 62, 453–471. ADVERTISERS Acknowledgments: We thank all contributors of NMR spectra for the training dataset. This project has received funding from the European Union’s Horizon 2020 research and innovation AUTHOR INDEX programme under the Marie Skłodowska-Curie grant agreement No 891690. PO010 POSTERS / BIOMOLECULES AND INTERACTIONS 163 NATHANIEL Z. HARDIN+1, NMR STUDIES ON METAL-CHELATED POLYMER NANODISCS USED VOJČ KOCMAN+2, IN COMBINATION WITH G-QUADRUPLEXES GIACOMO M. DI MAURO1, THIRUPATHI RAVULA1, There is considerable interest in the NMR community to speed up topologies. The T data that we gathered for the wtTel23 G-quadru-1 AYYALUSAMY data acquisition in NMR structural studies on biomolecules. One plex suggest that wtTel23 interacts with the nanodiscs by its groove RAMAMOORTHY1* way to achieve this is to significantly shorten the recycle delay in or loop regions and not through stacking on the nanodiscs by the 1Biophysics Program and Department of NMR experiments by speeding up the spin lattice relaxation time top or bottom G-quartet. Such a model of interactions is also sup-Chemistry, University of Michigan, Ann (T ) by applying paramagnetic relaxation enhancement (PRE). We ported by saturation transfer difference (STD) NMR experiments. Arbor, Michigan, USA 1 2National Institute of Chemistry, have focused on a modified poly-(styrene-co-maleic acid) polymer We show that to utilize the PRE effect we had to remove excess KCl Slovenian NMR Centre, Ljubljana, which forms nanodiscs with lipids and possesses the ability to salt from the G-quadruplex and nanodisc solution. We believe that Slovenia chelate Cu2+ ions [1]. Using inverse recovery experiments, we were the KCl salt reduces the interactions between the G-quadruplex and able to show that the T rates of protons for both polymer and lip-the nanodisc due to the salt charge screening between the DNA and 1 id-nanodisc components are significantly reduced, in some cases the lipid heads. even 7-fold, when the nanodiscs are Cu2+-chelated. This approach We believe that the described system could be especially valuable could potentially be very useful when structurally characterizing for NMR structural studies of large size RNA that exhibit very long lipid soluble molecules such as transmembrane proteins with NMR T values for protons and could enable multidimensional NMR ex-spectroscopy. Additionally, we show that Cu2+-chelated nanodiscs 1 perimental studies on membrane-associated peptides and proteins are also capable of decreasing the proton T values for a water-solu- 1 that may not be available in large quantity and/or are sensitive to ble biomolecule, the human telomere (wtTel23) DNA G-quadruplex. heat for long data acquisition [2]. This study also creates potential INTRODUCTION When comparing G-quadruplexes to double stranded DNA we see avenues to use the paramagnetic nature of the chelated polymer SPONSORS that their cores are comprised out of stacked G-quartets, arrange- nanodiscs for dynamic nuclear polarization (DNP) solid-state NMR PROGRAM ments of four guanine residues and that they form four grooves experiments to overcome the sensitivity issues in studying mem- PROGRAM — BY DAY instead of two. The guanines in G-quartets are connected by differ- brane proteins [3]. TOPICS ently structured loop regions that further diversify G-quadruplex PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES +These authors contributed equally to this work POSTERS References: [1] N. Z. Hardin, V. Kocman, G. M. Di Mauro, T. Ravula and A. Ramamoorthy, Angew. Chem. Int. Ed. 2019, 58, 17246-17250. [2] S. C. Keane, et. al., Science. 2015, 348, 917-921. [3] V. ADVERTISERS Kocman, G. M. Di Mauro, G. Veglia, A. Ramamoorthy, Solid State Nucl. Magn. Reson. 2019, 102, 36–46. AUTHOR INDEX Acknowledgments: This study was supported by funds from NIH (GM084018 to A.R.). PO011 POSTERS / BIOMOLECULES AND INTERACTIONS 164 MATIC KOVAČIČ1,2, UNIQUE STRUCTURAL FEATURES OF PYRENE- PETER PODBEVŠEK1,2, MODIFIED G-QUADRUPLEX REVEALED BY NMR HISAE TATEISHI-KARIMATA3, SHUNTARO TAKAHASHI3, G-quadruplexes are noncanonical secondary structures formed by guanine-rich nucleic acid strands that play roles NAOKI SUGIMOTO3,4, in various cellular functions. As independent aptamer molecules, G-quadruplexes are able to bind specific target JANEZ PLAVEC1,2,5 proteins and therefore represent potential drug candidates for the treatments of numerous diseases, most nota- 1Slovenian NMR Centre, National Institute of bly cancer. Small polyaromatic compounds can interact with solvent accessible DNA heterocyclic bases through Chemistry, Ljubljana, Slovenia 2 aromatic stacking, which leads to the stabilization of G-quadruplexes and subsequently to the regulation of their EN-FIST Centre of Excellence, Ljubljana, Slovenia functions. Alternatively, various chemical moieties and nucleotide analogs can be incorporated into G-quadruplex 3Frontier Institute for Biomolecular structure to fine-tune desirable properties of aptamers including structural stability, resistance against nuclease Engineering Research (FIBER), Konan degradation, and binding of target proteins. University, Kobe, Japan 4Graduate School of Frontiers of Innovative Research in Science and Technology (FIRST), Thrombin-binding aptamer (TBA) is a 15-mer 5’-d[GGTTGGTGTGGTTGG]-3’ DNA oligonucleotide, that folds into a Konan University, Kobe, Japan chair-type G-quadruplex capable of binding and inhibiting thrombin protease in the presence of sodium and po- 5Faculty of Chemistry and Chemical tassium cations [1,2]. In our study, TBA G-quadruplex served as a well-defined starting model which we modified Technology, University of Ljubljana, Ljubljana, Slovenia by substitutions of individual thymines with fluorescent Upy (5-(pyrene-1-yl-ethynyl)-dUMP) nucleotides [3]. Our results show that in the presence of potassium ions the individual replacements of T4, T9, and T13 with Upy nucleotides stabilize G-quadruplex and greatly increase its resistance against nuclease degradation while retaining the TBA fold. In the case of T9 substitution, a dynamic equilibrium between the unimolecular and bimolecular G-quadruplex structure was observed, the latter being comprised of two distinct G-quadruplex units. We believe that our INTRODUCTION approach could potentially be used for the optimization of various therapeutically important G-quadruplex-based SPONSORS aptamers. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] L. Bock, Nature. 1992, 355, 564-566. [2] A. Pica, FEBS J. 2013, 280, 6581-6588. [3] M. Kovačič, Nucleic Acids Res. 2020, 48, 3975-3986. PO012 POSTERS / BIOMOLECULES AND INTERACTIONS 165 MARTINA NMR INSIGHT INTO G-QUADRUPLEX FORMATION IN THE LENARČIČ ŽIVKOVIĆ1, REGULATORY REGION OF OSTEOPOROSIS-RELATED RANKL GENE JANEZ PLAVEC1,2,3 1 Pathogenic conditions that affect the skeleton, such as osteoporo- which is especially important for providing the optimized stacking Slovenian NMR Centre, National Institute of Chemistry, Ljubljana, sis, disrupt balanced bone remodeling by accelerating the differen- between G-quartets [4]. On the other hand, RAN4 forms a two-quar- Slovenia; tiation and maturation of osteoclasts. The pivotal regulator of os- tet structure stabilized by two base triads. G/A-to-T substitutions of 2Faculty of Chemistry and teoclast activity is the receptor activator of NF-κB (RANK) expressed the residues from base triads uncovered the critical role of A5 for Chemical Technology, University of Ljubljana, Ljubljana, Slovenia; by osteoclasts that interacts with RANK ligand (RANKL) formed by the formation of a distinct two-quartet topology; A5-to-T5 substi-3EN-FIST, Centre of Excellence, osteoblasts. The interaction initiates a cascade of intracellular sig- tution switched the two-quartet structure into three-quartet (3+1) Ljubljana, Slovenia naling events and promotes bone resorption that in the case of ex- hybrid G-quadruplex [5]. cessive activity leads to osteoporosis [1]. One of the potential ways of To test whether the presence of a G-rich sequence able to form gene regulation on transcriptional level is facilitation or inhibition G-quadruplexes can affect the gene expression levels, a fragment of of transcription by uncanonical secondary structures formation in the human RANKL proximal promoter region was cloned into the the G-rich fragments of promoter regions [2,3]. A G-rich region in luciferase report vector (collaboration with Faculty of Pharmacy, the RANKL gene proximal promoter sequence with the potential to University of Ljubljana, Slovenia). Different mutations in the G-rich adopt G-quadruplex structures was identified using bioinformatics. region were analyzed, including RAN1, RAN4, and deletion of the A 20-nt long sequence RANwt, d(GGGGAGGGAGCGGGAGAGGG), entire G-rich sequence. The latter resulted in increased expression folded into diverse structures, presumably due to four consecutive levels in HeLa and A549 cell lines, suggesting that the presence of guanines in the first G-tract. Individual substitutions of G1 and G4 the G-rich sequence causes repression of RANKL promoter activity. INTRODUCTION with thymine led to single, although completely different G-quadru- Our results, to the best of our knowledge, are the first to suggest pos- SPONSORS plex conformations RAN1 and RAN4, respectively. High-resolution sible regulation mechanisms of the RANK/RANKL pathway by the PROGRAM NMR structure showed that RAN1 folds into a parallel G-quadruplex formation of noncanonical DNA structures in the regulatory region PROGRAM — BY DAY with three G-quartets connected by 3 propeller loops, and a bulge. of the RANKL gene. TOPICS Residues from the G-A bulge adopt a pseudo-loop conformation, PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Rao et al. , Trends Cell Biol. 2018, 28, 213-223; [2] Bochman et al. , Nat. Rev. Genet. 2012, 13, 770-780; [3] Holder and Hartig, Chem. Biol. 2014, 21, 1511-1521; [4] Lenarčič Živković, POSTERS Rozman, Plavec, Molecules 2020, 25 (20): 4867; [5] Lenarčič Živković, Rozman, Plavec, Angew. Chem. In. Ed. 2018, 57, 15395-15399. ADVERTISERS Acknowledgments: The authors acknowledge the financial support from Slovenian Research Agency (research core funding No. P1-242 and J1-1704) and the CERIC-ERIC Consortium for AUTHOR INDEX access to experimental facilities. PO013 POSTERS / BIOMOLECULES AND INTERACTIONS 166 TEVŽ LEVSTEK1,2, STUDY OF STRUCTURAL MOTIFS ADOPTED BY MARIA TOPLISHEK 1,3, RNA SEQUENCES WITH ADENINE, GUANINE, AND JANEZ PLAVEC1, 2, 3 1National Institute of Chemistry, CYTOSINE REPEATS Slovenian NMR Centre, Ljubljana, Slovenia Recently, a new DNA tetrahelical structure was discovered in our In the following steps, we studied sequences without G-tracts de-2University of Ljubljana, Faculty department, formed by sequences with AGCGA repeats, which signed in such a way that would enable the formation of tetrahelical of Chemistry and Chemical Technology, Ljubljana, Slovenia consists of mixed GAGA and GCGC quartets [1]. This type of motifs secondary structures involving mixed quartets of adenine, guanine, 3EN-FIST Centre of Excellence, has never been described for RNA, therefore the study of structures and cytosine. Initially different types of sequences were tested, how-Ljubljana, Slovenia adopted by a family of RNA oligonucleotide sequences containing ever, our efforts were mostly focused on sequences with the propen-mixed adenine, guanine, and cytosine repeats is important due to sity of folding into parallel type tetrahelical structures. the fact that RNA secondary structures are often involved in vari- The dynamic nature of studied oligonucleotides manifested as an ous regulatory processes including protein expression. Also, such equilibrium between multiple species including hairpin structures sequences are relatively common in vertebrate (including human) made the investigation challenging. Nevertheless, a few intriguing transcriptomes which indicates their biological relevance. trends were found. Namely, some of the designed sequences lead We initiated a study on RNA oligonucleotides with tandem AGCGA to the formation of different structures depending on solution repeats. RNA oligonucleotides demonstrated drastically different conditions. We also observed folding topologies which are charac-behavior in comparison to DNA counterparts. They showed a pref- terized by stabilization of non-canonical base pairs in addition to erence for G-quadruplex formation rather than AGCGA-quadru- Watson-Crick pairs, which is unlikely for the hairpin. NMR signals INTRODUCTION plex. Not too surprisingly, as the 2′-OH hydroxyl groups in the RNA of studied RNA oligonucleotides allowed us to propose a partially SPONSORS quadruplex play a significant role in redefining hydration structure symmetrical tetrahelical structure. PROGRAM in the grooves and the hydrogen-bonding networks. This structural PROGRAM — BY DAY change for example increases the stability of RNA telomeric qua- TOPICS druplexes over DNA ones [2]. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] V. Kocman, J. Plavec, Nature Commun. 2017, 8, 15355. [2] G.W. Collie, S.M. Haider, S. Neidle, G.N. Parkinson, Nucleic Acids Res. 2010, 38, 5569-5580. PO014 POSTERS / BIOMOLECULES AND INTERACTIONS 167 MAJA MARUŠIČ1, TEMPERATURE, CATION, AND STRAND JANEZ PLAVEC1,2,3 CONCENTRATION CONTROLLABLE DNA SWITCH 1 Slovenian NMR Centre, National Institute of Chemistry, Ljubljana, Nucleic acids are carriers of molecular information in biology. But they are more than that - as biopolymers, they Slovenia 2University of Ljubljana, Faculty have exciting physicochemical properties, which can be rationally influenced by the base sequence. Additionally, of Chemistry and Chemical some non-double-stranded structural variations adopted by oligonucleotides with various repetitive sequences ex-Technology, Ljubljana, Slovenia 3 pand the potential of nucleic acids to be used for building blocks for nanotechnology, to self-assemble into molec-EN-FIST Centre of Excellence, Ljubljana, Slovenia. ular nanostructures, and to be used as a material for machine-like nanodevices. To be assembled and to change in a controllable manner, nucleic acids were designed to respond with structural change to external stimuli, such as photons, temperature, pressure, magnetic or electric fields, or altered chemical environments. Nucleic acid systems that respond to several stimuli in a different way enable the design of multifunctional devices [1]. We describe a short, 17 nucleotides long DNA switch that is capable of adopting six different structures in response to temperature, cation, and DNA strand concentration. NMR analysis identified the type of structures present at a certain combination of external stimuli and conditions at which they interconvert. Change in distance between oligonucleotide ends for each of the structures is the basis for the design of a reporter signal and the development of multifunctional nanodevices. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] Krishnan, Y. & Simmel, F. C. 2011, Ange. Chem. Int. Ed. 50, 3124–3156 (2011). PO015 POSTERS / BIOMOLECULES AND INTERACTIONS 168 NATAŠA MEDVED1, METHYLATION OF CYTOSINE IN THE PROMOTER REGION OF BCL2 MIRKO CEVEC1, DOES NOT IMPACT STRUCTURE JANEZ PLAVEC1,2,3 1National Institute of Chemistry, Methylation of cytosine in the CpG dinucleotide step by the enzyme into the groove that is close to the H1’/H4’ side of the G3 sugar. Since Slovenian NMR Centre, Ljubljana, DNA methyltransferase (DNMT) occurs at the C5 position of the the NOEs that position mC4 in the groove are weaker compared to Slovenia 2EN-FIST Center of Excellence, cytosine to form 5-methylcytosine (mC). The methyl group on cytosine the parent Bcl2Mid, we consider the positioning of mC4 inside the Ljubljana, Slovenia alters both stereoelectronic and hydrogen bond properties. However, groove as less favorable. 3University of Ljubljana, Faculty mC can still base pair with a guanine residue on the complementary of Chemistry and Chemical In the case of mC6 substituted oligonucleotide, we observed small Technology, Ljubljana, Slovenia strand, leaving the DNA coding capacity unaltered. But the introduc- differences in distances between the mC6 and adjacent guanines in tion of epigenetic modification into the gene promoter region could the upper G-quartet. affect the DNA local structure and its stability [1]. No detectable differences compared to the parent Bcl2Mid were ob- We studied the impact of mC on the G-quadruplex structure using served in the NOESY spectrum in the case of mC at position C20. a model sequence (5’-GGGCGCGGGAGGAATTGGGCGGG-3’) from the Bcl2Mid promoter region. Bcl2Mid oligonucleotide has been Thermal stabilities of the parent G-quadruplex and mC analogs shown to form an intramolecular (3+1) G-quadruplex structure in were comparable (±1°C). the presence of K+ ions [2]. Special attention was paid to the first CD spectra of Bcl2Mid G-quadruplex and analogs with mC in the loop region of Bcl2Mid (C4-G5-C6) which defines and stabilizes the presence of K+ ions were very similar, with a negative peak near 240 Bcl2Mid structure due to its specific way of interaction with the core nm and two positive peaks at 270 and 295 nm, which is in agree- G-quartets. INTRODUCTION ment with the (3+1) hybrid G-quadruplex structure. SPONSORS We introduced mC at positions 4, 6, and 20 of the parent Bcl2Mid The structural changes caused by the presence of mC at positions PROGRAM oligonucleotide. In the respective 1D 1H NMR spectra we observed 4, 6, and 20 are small and limited to local perturbations. Based on PROGRAM — BY DAY twelve well-resolved imino proton resonances located between 10.5 2D NOESY structural data we can infer that the structural impact of TOPICS and 12.0 ppm that clearly indicate the formation of a G-quadruplex methylation of cytosine in the CpG dinucleotide steps of Bcl2Mid PRIZE LECTURES structure with three G-quartets. oligonucleotide is minimal and causes no significant structure PLENARY LECTURES C4 adopts an unusual orientation in the parent Bcl2Mid structure. rearrangements. TUTORIAL LECTURE It isn’t stacked over the G3·G23·G19·G7 quartet but is positioned INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] T. Pfaffender, B. Hackner, M. Truß, Angew Chem Int Ed Engl. 2011, 50, 7008-7012; [2] J. Dai, D. Chen, R.A. Jones, L.H.Hurley, D.Yang, Nucleic Acids Res. 2006, 34, 5133-5144. AUTHOR INDEX Acknowledgments: This project was supported by Slovenian Research Agency [ARRS, grant P1-0242]. PO016 POSTERS / BIOMOLECULES AND INTERACTIONS 169 DEEPRAJ NEGI, MAPPING MOLECULAR INTERACTIONS THAT DRIVE RAJESH KUMAR REDDY G-QUADRUPLEX CONFORMATION AND STABILITY SANNAPUREDDI, BHARATHWAJ G-Quadruplexes (G4s) are non-canonical secondary structures of nucleic acid formed by guanine rich sequences. SATHYAMOORTHY G4s are the four stranded DNA structures comprising four guanines in one plane known as G-quartet, for three Department of Chemistry, Indian planes G4s each G strand contains at least three G-residue separated by loops. G-quartet which is formed by the Institute of Science Education and interaction of guanine from each strand is stabilized by Hoogsteen hydrogen bonds, stacking of quartet and central Research Bhopal, Bhauri, Madhya Pradesh, India counter ion typically monovalent cations. Interestingly, G4 structures are polymorphic in nature and depending upon the cation present and the nucleotides in the loops are evidently found in different topologies like parallel, hybrid (3+1), antiparallel (2+2) and antiparallel. G4s are known to be present in various regions of the human genome such as telomere, promoter and 5’-UTRs [1]. They have gained enormous attention due to their implication in several crucial biological processes like replication, transcription, translation and telomere maintenance [2] which makes G4s a potential drug target [3]. This study focuses on mapping molecular interactions that drive G4s conformations and stability. To decipher the conformational complexity and the thermodynamic stability of G4s we have chosen the model system TTGGGQ: 5’-AAGGG(TTGGG)3AA-3’ which forms parallel conformation in K+ and Hybrid (3+1) in Na+ conditions [4]. The TTG- GGQ sequence has been modified by substituting each guanine with inosine to reduce one hydrogen bond at a time to check the role each guanine plays in the stability. CD spectroscopy has been used to characterize different topologies of G4s [5], herein we observed the biphasic behavior for TTGGGQ and inosine modified sequences in K+ INTRODUCTION condition with a reduction in thermal stability(T ) for the latter case. With NMR spectroscopy, we observed that in m SPONSORS K+ condition inosine modification favors a single as well as multiple conformations (in few cases), giving the valida- PROGRAM tion of some crucial molecular interactions which are responsible to drive a particular G4 conformation. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Antonio D. M. et al, Nat Chem. 2020, 12, 832-837. [2] Varshney D. et al, Nat Rev Mol Cell Biol. 2020, 21, 459-474. [3] Jamroskovic J. et al, J Am Chem AUTHOR INDEX Soc. 2020, 142, 2876-2888. [4] Sannapureddi R. et al, J. Phys. Chem. Lett. 2020, 11, 10016-10022. [5] Guerra R. et al, Angew. Chem. Int. Ed. 2018, 57, 7171-7175. PO017 POSTERS / BIOMOLECULES AND INTERACTIONS 170 ALEŠ NOVOTNÝ1, STRUCTURAL STUDY OF CGAG-RICH SEQUENCE VOJČ KOCMAN1, FROM THE PROMOTER OF AUTS2 GENE INVOLVED IN JANEZ PLAVEC1,2,3 1National Institute of Chemistry, NEURODEVELOPMENT Slovenian NMR Centre, Ljubljana, Slovenia The human AUTS2 gene has been shown to importantly influence brain development by controlling the number of 2EN-FIST Centre of Excellence, neurons, affecting the structure of neurons by promoting the growth of axons and dendrites as well as regulating Ljubljana, Slovenia 3Faculty of Chemistry and neuronal migration [1]. AUTS2 is also a key regulator of transcription of multiple genes crucial for brain devel-Chemical Technology, University opment and misregulations in AUTS2 expression have been correlated with the occurrence of autism spectrum of Ljubljana, Ljubljana, Slovenia disorders [2,3]. A CGAG-rich sequence (A38) capable of adopting a non-canonical secondary structure was found in the promoter region of AUTS2 approximately 150 bases upstream of the transcription start site. Using NMR complemented by UV/Vis spectroscopy and gel electrophoresis we show that A38 adopts a stable non-canonical hairpin structure composed of six GC, four GA base pairs, and a 14-nucleotide loop. Furthermore, the A38 sequence is rich in CpG dinucleotides which are known hotspots for epigenetic modifica- tions. The most abundant epigenetic modification in the human genome is 5-methylcytosine that is involved in gene silencing [4]. The demethylation process can start by deamination of 5-methylcytosine which yields thymine now associated in a G:T mismatch. This mismatch is readily recognized by thymine DNA glycosylase and converted to cytosine via a base excision repair pathway [5]. We show that the incorporation of a single C-to-T mutation in cer- INTRODUCTION tain positions of the A38 sequence alters the structure of the hairpin. This change significantly affects the structure SPONSORS of the loop region and can be understood as four-nucleotide slippage compared to the native sequence. The for- PROGRAM mation of different structures based on epigenetic modifications, or their repair pathway intermediates, may play PROGRAM — BY DAY important role in the regulation of AUTS2 expression. Additionally, the high-resolution structures will help expand TOPICS our knowledge of structures formed by repetitive sequence motifs. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Oksenberg, N., Stevison, L., Wall, J. D., Ahituv, N. PLoS Genet. 2013, 9. [2] Pang, W., Yi, X., Li, L., Liu, L., Xiang, W., Xiao, L. Front. Psychiatry 2021, POSTERS 12. [3] Hori, K. et al. iScience 2020, 23. [4] Jang, H. S., Shin, W. J., Lee, J. E., Do, J. T. Genes 2017, 8. [5] Cortellino, S. et al. Cell 2011, 146. ADVERTISERS Acknowledgments: Authors acknowledge the financial support of the Slovenian Research Agency [ARRS, grant P1-0242, A.N. for Young AUTHOR INDEX researcher grant]. PO018 POSTERS / BIOMOLECULES AND INTERACTIONS 171 DAŠA PAVC1,2, ROLE OF CATION NATURE AND GC ENDS IN THE BAIFAN WANG1, ASSEMBLY OF DNA G-QUADRUPLEXES LEA SPINDLER3,4, IRENA DREVENŠEK-OLENIK4,5, Guanine-rich DNA oligonucleotides can adopt non-canonical, four-stranded secondary structures, termed G-qua- JANEZ PLAVEC1,2,6 AND druplexes. Their main building block is a G-quartet, which is formed by four guanine residues in planar arrangement PRIMOŽ ŠKET1 held together by eight Hoogsteen-type hydrogen bonds. The formation of G-quadruplexes requires the presence of 1National Institute of Chemistry, Slovenian NMR cations. In fact, cation nature is one of the major factors contributing to the structural diversity of G-quadruplexes. Centre, Ljubljana, Slovenia 2 Their self-assembling ability as well as programmable control of their shape and size make them attractive candi-University of Ljubljana, Faculty of Chemistry and Chemical Technology, Ljubljana, Slovenia dates for nanotechnological applications. One of the proposed ways for programming self-assembly is by designing 3University of Maribor, Faculty of Mechanical G-quadruplex forming DNA sequences with complementary GC ends, which would act as ‘sticky ends’ and form Engineering, Maribor, Slovenia 4 linkages between two successive G-quadruplexes via inter quadruplex GCGC-quartet formation [1, 2]. Jozef Stefan Institute, Department of Complex Matter, Ljubljana, Slovenia 5University of Ljubljana, Faculty of Mathematics In the present NMR study, we analyzed the influence of GC ends within oligonucleotides GCn and GCnCG, where n = and Physics, Ljubljana, Slovenia G AG AG on resulting G-quadruplex structures. Additionally, we evaluated the effect of different monovalent cat-2 4 2 6EN-FIST Center of Excellence, Ljubljana, ions on the folding of GCn and GCnCG G-quadruplexes. Na+, 15NH + and K+ ions promote the formation of symmetric, 4 Slovenia dimeric G-quadruplexes, which can be considered, as structures composed of two blocks the 5’-antiparallel and the 3’-parallel block. Although the global fold of G-quadruplexes, which includes A(GGGG)A hexad sandwiched between two G-quartets is preserved, some structural elements were cation dependent. Furthermore, 15NH + and K+ 4 ions promote dimerization of GCn G-quadruplexes through 3’-3’ stacking interactions of terminal G-quartets. Such stacking is precluded by 3’-GC ends in the case of GCnCG G-quadruplexes [3]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] T. Ilc, P. Šket, J. Plavec, M. Webba da Silva, I. Drevenšek Olenik, L. Spindler, J. Phys. Chem. C. 2013, 117, 23208-23215. [2] T. Troha, I. Drevenšek- POSTERS Olenik, M. Webba da Silva, L. Spindler, Langmuir. 2016, 32, 7056-7063. [3] D. Pavc, B. Wang, L. Spindler, I. Drevenšek-Olenik, J. Plavec, P. Šket, Nucleic Acid Res. ADVERTISERS 2020, 48, 2749-2761. AUTHOR INDEX Acknowledgment: This work was supported by the Slovenian Research Agency (ARRS, grants P1-0242 and J1-7108). PO019 POSTERS / BIOMOLECULES AND INTERACTIONS 172 KATEŘINA PETERKOVÁ1,2,3, c-kit2 G-QUADRUPLEX STABILIZED VIA A COVALENT IVO DURNÍK2,4, PROBE: EXPLORING G-QUARTET ASYMMETRY RADEK MAREK2,4,5, JANEZ PLAVEC1,3,6 AND The KIT receptor is a transmembrane protein that participates in a variety of physiological processes [1]. Due to its PETER PODBEVŠEK1 role in the pathogenesis of cancer, KIT is an attractive target for anti-cancer treatment [2]. The human KIT proto-on-1 National Institute of Chemistry, Slovenian cogene promoter contains three G-rich regions, c-kit1, kit*, and c-kit2, which are capable of folding into G-quadru-NMR Centre, Ljubljana, Slovenia 2 plexes. Importantly, the promoter segment comprising kit* and c-kit2 contains a putative binding site for the Sp1 Masaryk University, Faculty of Science, National Centre for Biomolecular Research, transcription factor, which can (apart from binding to a double-stranded consensus motif) bind to a G-quadruplex Brno, Czechia [3]. Considering that Sp1 binding is critical for the activity of the human KIT promoter [4], highly stable G-rich oligo-3 University of Ljubljana, Faculty of nucleotides mimicking G-quadruplexes from KIT could be used as decoys to sequester these proteins and modulate Chemistry and Chemical Technology, Ljubljana, Slovenia KIT expression. 4 Masaryk University, CEITEC-Central European Institute of Technology, Brno, Polyaromatic moieties can be employed for modulating G-quadruplex properties via their stacking with G-quartets. Czechia In this study, we focused on G12T/G21T mutant of the genomic c-kit2 sequence forming a monomeric three-quar- 5 Masaryk University, Faculty of Science, Department of Chemistry, Brno, Czechia tet G-quadruplex, as a representative of parallel G-quadruplex structures found in human promoter regions. We 6 EN-FIST Centre of Excellence, Ljubljana, showed that individual incorporation of Upy (5-(1-pyrenylethynyl)-2’-deoxyuridine) in the pentaloop of c-kit2 caused Slovenia structural polymorphism and in some cases also destabilization. On the other hand, the introduction of pyrene moieties to an individual or both termini of the c-kit2 sequence resulted in highly stable G-quadruplex structures. Although the parent parallel fold remained unchanged despite the terminal substitutions, a detailed analysis revealed major differences in structural dynamics of Upy between the two terminal analogues. We believe that the con- INTRODUCTION trast between structural dynamics of Upy1 and Upy21 might stem from an intrinsic asymmetry of c-kit2 G-quartets. SPONSORS This way Upy acts as a probe for local G-quadruplex dynamics, which is true especially for c-kit2, where outer G-quar- PROGRAM tets are exposed and Upy interactions with propeller loops are minimized. This is a vice-versa effect to the binding of PROGRAM — BY DAY ligands comprised of unfused aromatic rings to G-quadruplexes, where ligand planarity is key for efficient stacking. TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] C. E. Edling, B. Hallberg, Int. J. Biochem. Cell Biol. 2007, 39, 1995–1998. [2] L. K. Ashman, R. Griffith, Expert Opin. Investig. Drugs. 2013, 22, TUTORIAL LECTURE 103–115. [3] E. A. Raiber, R. Kranaster, E. Lam, M. Nikan; S. Balasubramanian, Nucleic Acids Res. 2012, 40, 1499–1508. [4] G. H. Park, H. K. Plummer, G. W. Krystal, Blood. 1998, 92, 4138–4149. INVITED AND PROMOTED LECTURES Acknowledgments: This work was supported by the European Programme H2020 MSCA ITN [grant number 765266-LightDyNAmics project]. The authors POSTERS also acknowledge financial support from the Slovenian Research Agency [grants P1-0242 and J1-1704]. The Ministry of Education, Youth and Sports of ADVERTISERS the Czech Republic (MEYS CR) is acknowledged for its support of access to research infrastructure (CEITEC 2020 LQ1601, CIISB: LM2018127, e-INFRA CZ: AUTHOR INDEX LM2018140). PO020 POSTERS / BIOMOLECULES AND INTERACTIONS 173 STASĖ BIELSKUTĖ1, THE EFFECT OF 8-OXOGUANINE ON G-QUADRUPLEX STRUCTURE JANEZ PLAVEC1,2,3, PETER PODBEVŠEK1 Reactive oxygen species (ROS) are a byproduct of aerobic cellular replacing guanines with different glycosidic conformations. Ac-1 metabolism in all living organisms. The inability to neutralize ex- commodation of oxoG at sites in syn or anti in non-substituted hTel National Institute of Chemistry, Slovenian NMR Centre, cessive ROS results in oxidative stress causing many types of DNA G-quadruplex requires a minor structural rearrangement or a major Ljubljana, Slovenia lesions, to which cells respond by activating relevant DNA repair conformation shift, respectively. The system responds by retaining 2EN-FIST Center of Excellence, pathways or apoptosis if repair is unsuccessful. Among the four or switching to a fold where oxoG is in syn conformation. Important-Ljubljana, Slovenia 3 Faculty of Chemistry and DNA nucleobases, guanine (G) has the lowest redox potential and is ly, the two G-quadruplex structures containing oxoG are still stable at Chemical Technology, University therefore most prone to oxidation. Furthermore, guanines in tracts physiological temperatures and should be considered detrimental of Ljubljana, Ljubljana, Slovenia are more susceptible to oxidation than isolated guanines. in higher-order telomere structures [1]. G-rich regions are found at telomeric ends of chromosomes and fold Human promoters are also enriched in G-quadruplex forming se-into G-quadruplex structures. Four repeats of the human telomer- quences. Misregulation of BCL2 expression has been observed in ic sequence (hTel) form three stacked G-quartet planes with mixed many diseases and is associated with cellular exposure to reactive parallel/antiparallel G-tract directionalities. We have individual- oxygen species. A region upstream of the P1 promoter in the human ly probed all guanine positions in hTel by substituting them with BCL2 gene plays a major role in regulating transcription. A similar 8-oxo-7,8-dihydroguanine (oxoG), a common oxidation product of strategy was used to simulate oxidative events within a long G-tract guanine. Due to oxoG’s distinct hydrogen bonding properties, a loss by oxoG substitutions. Surprisingly, oxoG at a specific position within of G-quadruplex structure was observed for most oligonucleotides a 25-nt construct boosts thermal stability of the resulting G-qua-containing oxidative lesions. However, some positions in the hTel druplex. This is achieved by distinct hydrogen bonding properties INTRODUCTION sequence can tolerate substitutions with oxoG. Two positions, which of oxoG, which facilitate the formation of an antiparallel basket-type SPONSORS adopt anti and syn glycosidic conformations in the parent hTel G-quadruplex with a three G-quartet core and a G·oxoG·C base tri- PROGRAM G-quadruplex, respectively, were selected for the further analysis ad (PDB ID: 6ZX6). While oxoG has previously been considered det- PROGRAM — BY DAY and determination of high-resolution structures by solution-state rimental for G-quadruplex formation, its stabilizing effect within TOPICS NMR (PDB IDs: 6IA0 and 6IA4). However, due to oxoG’s preference a promoter suggests a potential novel regulatory role of oxidative PRIZE LECTURES for the syn conformation, distinct responses were observed upon stress in general and specifically in BCL2 gene transcription [2]. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] S. Bielskute, J. Plavec, P. Podbevšek, J. Am. Chem. Soc. 2019, 141, 2594-2603. [2] S. Bielskute, J. Plavec, P. Podbevšek, Nucleic Acids Res. 2021, 49, 2346- AUTHOR INDEX 2356. PO021 POSTERS / BIOMOLECULES AND INTERACTIONS 174 ADITYA POKHARNA, LOCAL THERMODYNAMICAL INVESTIGATION OF JULIEN ORTS, Pin1 WW-DOMAIN USING eNOEs ROLAND RIEK ETH Zurich, Department The traditional way of determining the structure of bio-molecules domain) at local level and expand the utility of eNOE toolkit. The of Chemistry and Applied using solution-state NMR involves using a large set of NOE rates de- 34 residue WW domain used in this study is the N-terminal part of Biosciences, Zurich, Switzerland rived from NOESY spectra and converting them to inter-proton dis- Pin1 and folds into the three-stranded anti-parallel beta-sheets, typ- tances. The biggest limitation of this method is that we are only able ical of WW-domains. Over the years, multiple kinetic and thermo- to semi-quantitatively determine the cross-relaxation rates from dynamic studies of WW-domain have shown that it displays a strict the spectra and consequently the distances that we obtain only two-state behavior [2]. work as upper-limits of the actual distances with huge tolerances. We extracted the structural information of the protein for a wide The main reason behind this is spin diffusion, the higher-order temperature range (278K-303K) and tried to model the evolution magnetization transfer to the spins outside of the two spins being of the obtained rates for every spin pair with increasing tempera-studied. ture under a two-state thermodynamics paradigm. This approach One of the methods to provide accurate values of rates and distanc- enabled us to extract parameters like enthalpy, ΔH, heat capacity, es and in turn more precise structures is exact NOEs (eNOEs) de- ΔC , and temperature, T (where ΔG = 0), for proton pairs spread p t veloped by Vogeli et al. [1]. This method utilizes a hybrid approach across the breadth of protein which were inaccessible by other ap-where we use an X-ray or a preliminary NMR structure and subject it proaches, as they only gave us information at the global level. to full-matrix formalism to determine the ratio of magnetization The mean value of parameters T, ΔH and ΔC is 289.03 K, 5.71*10^4 transfer between the two spins in question with and without the t t p,t INTRODUCTION J/mol and -2.14*10^3 J/Kmol, respectively and the standard devia- influence of the neighboring spins. This ratio, called the correction SPONSORS tion is 9.83 K, 1.51*10^4 J/mol and 4.80*10^3 J/Kmol, respectively. factor is then applied to all experimentally determined NOE build- PROGRAM The values of T and ΔH seem to be constrained within a narrow ups to gain precise and unambiguous values of relaxation rates and t t PROGRAM — BY DAY range and have relatively low variance. This hints towards some distances, with both upper and lower limits. TOPICS kind of correlated transition state at ~ 290K, much below the PRIZE LECTURES In our work, we use the eNOE rates obtained through this method to melting temperature (~ 320K). This is in line with the two-state PLENARY LECTURES study thermodynamics of protein (S18N/W34F mutant of Pin1-WW structure we obtained with our protein allostery calculations [3]. TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] B. Vogeli, S. Kazemi, P. Guntert, R. Riek, Nat. Struct. Mol. Biol. 2012, 19(10):1053–1057. [2] J. Crane, E. Koepf, J. Kelly, and M. Gruebele. J. Mol. Biol. 2000, 298(2):283–292. [3] D. AUTHOR INDEX Strotz, J. Orts, H. Kadavath, R. Riek et al. Angew. Chem. Int. Ed. 2020, 59 (49). PO022 POSTERS / BIOMOLECULES AND INTERACTIONS 175 LETIZIA PONTORIERO, ZOOMING ON THE INTERACTION BETWEEN MARCO SCHIAVINA, a-SYNUCLEIN AND CALCIUM IONS APPROACHING MARIA GRAZIA MURRALI, PHYSIOLOGICAL CONDITIONS ROBERTA PIERATTELLI, ISABELLA C. FELLI Intrinsically disordered proteins (IDPs) and protein regions (IDRs) lack a stable 3D structure but are nevertheless functional thanks to their dynamic properties [1]. Their fine-tuned features are expected to be modulated by side-Magnetic Resonance Center (CERM) and Department of Chemistry “Ugo chains as well as local solvent exposure [2] so it is crucial to characterize them with atomic-resolution approaches. Schiff”, Here we propose a 13C based strategy to provide a unique tool to investigate IDPs behavior in different experimental University of Florence, Sesto conditions relevant for their physiological function. A set of carbonyl carbon direct detected NMR experiments was Fiorentino, Florence, Italy. implemented to monitor the IDPs/IDRs both from the backbone and side chains point of view. The latter is seldom studied in highly flexible and disordered proteins because of extensive signal overlap [3]. In addition, a novel pulse sequence based on the CON experiment was designed to achieve information of amide proton exchange with the solvent (DeCON). This set of experiments was used to obtain a fingerprint of a-synuclein that is fully disordered in its native conditions and to study the interplay between this IDP and Ca2+ ions [4], zooming into the metal ion coordination sphere and revealing the motifs involved in the interaction [5]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] V. N. Uversky, Int. J. Biochem. Cell Biol. 2011 43, 1090–1103. [2] V.N. Uversky, Front. Mol. Biosci. 2014 1, 1–24. [3] I.C. Felli, R. Pierattelli, Intrinsically INVITED AND PROMOTED LECTURES Disordered Proteins Studied by NMR Spectroscopy, Springer, 2015. [4] A. Binolfi, R. M. Rasia, C. W. Bertoncini, M. Ceolin, M. Zweckstetter, C. Griesinger, T. POSTERS M. Jovin, C. O. Fernández, J. Am. Chem. Soc. 2006, 128, 9893–9901. [5] L. Pontoriero, M. Schiavina, M. G. Murrali, R. Pierattelli, I. C. Felli, Angew. Chem. Int. Ed. ADVERTISERS 2020, 59, 18537– 18545. AUTHOR INDEX Acknowledgments: CERM/CIRMMP center of Instruct-ERIC, Fondazione CR Firenze. PO023 POSTERS / BIOMOLECULES AND INTERACTIONS 176 ADRIANA RATHNER1, BUSY BAZAAR: WHAT HAS NMR TOLD US ABOUT MICHAL KAMENICKY2, EXTRINSIC PROTEINS OF PHOTOSYSTEM II? PETR RATHNER1, 3, VITUS STEMMER2, Psb proteins of Photosystem II are a class of extrinsic proteins that adjoin the oxygen evolving center from the cyto-NORBERT MÜLLER1, 2, 4 solic side. They act as gatekeepers who maintain proper conditions for the water splitting reaction which ultimately 1JKU Linz, Institute of Inorganic leads to the release of molecular oxygen in higher plants [1]. This is achieved by fine tuning the concentrations of Chemistry, Linz, Austria 2 ions present in the photosynthetic machinery via protein-protein and protein-ion interactions. The exact function-JKU Linz, Institute of Organic Chemistry, Linz, Austria al mechanisms of this protein cohort have not yet been described in detail since all of these proteins contain a large 3University of Vienna, Institute of degree of disordered regions which so far impeded the resolution of complete proteins by means of X-ray crystal-Analytical Chemistry, Vienna, Austria 4 lography or cryo-electron microscopy [1, 2]. University of South Bohemia, Faculty of Science, Ceske Budejovice, Czechia We have studied 3 main extrinsic proteins – PsbP (23 kDa), PsbQ (16 kDa) and PsbO (33 kDa) as well as the extrinsic domain of the larger membrane protein, CP43 (CP43ext., 12 kDa). Using solution NMR we have been able to determine three dimensional structures of PsbP and PsbQ in solution including their highly flexible regions. We have obtained partial resonance assignment and relaxation data of PsbO. Recently, we have initiated the NMR characterization of the extrinsic domain of CP43 which is in spatial proximity to all three extrinsic proteins. The most interesting information about detailed interplay of those proteins and their dependence on the presence of metal cations has been assessed using a combination of techniques, such as NMR titration and chemical exchange saturation transfer (CEST), biolayer interferometry (BLI) and microscale thermophoresis (MST). INTRODUCTION The metal binding sites we found in the solution differ substantially from the ones previously reported in crystal- SPONSORS lographic structures of isolated higher plants of PsbP and PsbQ [3, 4]. All of the protein interaction sites (with other PROGRAM proteins as well as the cations) included dynamic parts which suggest the so far unrecognized functional impor- PROGRAM — BY DAY tance of these regions. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] J. L. Roose, L. K. Frankel, M. P. Mummadisetti, T. M. Bricker, Planta. 2016, 243, 889-908. [2] X. Wei, X. Su, P. Cao et al., Nature. 2016, 534, 69-74. INVITED AND PROMOTED LECTURES [3] V. Kopecky Jr., J. Kohoutova, M. Lapkouski et al., PLoS ONE. 2012. [4] M. Balsera, J. B. Arellano, J. L. Revuelta et al., J Mol Biol. 2005, 350, 1051-1060. POSTERS Acknowledgments: NMR experiments were performed at 700 MHz Bruker AVANCE III (cryogenically cooled probe) at the NMR laboratory of the Upper ADVERTISERS Austrian - South Bohemian Research Infrastructure Center in Linz, co-financed by the European Union in the context of the project “RERI-uasb”, EFRE RU2- AUTHOR INDEX EU-124/100-2010 (ETC Austria-Czech Republic 2007-2013, project M00146). PO024 LOREM / POSTERS / BIOMOLECULES AND INTERACTIONS 177 RAJESH KUMAR REDDY CHEMICAL SHIFTS AND MACHINE LEARNING BASED SANNAPUREDDI, METHODOLOGY TOWARDS CHARACTERIZATION OF DNA MANISH KUMAR MOHANTY, ANOOP KUMAR GAUTAM, G-QUADRUPLEXES BHARATHWAJ NMR based structural genomics has immensely contributed to the glycosyl dihedral angles ( viz. “syn” and “anti”). From BMRB search, SATHYAMOOTHY field of structural biology. Tertiary structure modelling has seen a we observed that 1H-H8 CS aids in the dihedral angle distribution, Department of Chemistry, Indian Institute paradigmatic shift with the development of chemical shifts (CS) albeit with a lower level of accuracy due to significant overlap of of Science Education and Research Bhopal, based methodology. Similar protocols for nucleic acids are in their chemical shift distributions. Due to the dearth of 13C/15N data, 18 Madhya Pradesh, India infancy, with 1H CS based structural characterization for RNA be- different sequences were prepared (natural isotopic abundance) for ing the most recent. We envisioned that similar protocols for DNA which PDB structural models are available.[1] 2D 13C-1H and 15N-1H G-quadruplexes (G4) would be immensely helpful, given their di- correlation data were obtained for samples (concentrations 0.8-3 verse roles performed within the cell and also their applications mM) within a short period (8-30 hours/sample) to build the chem-across the fields of Chemistry and Biology. ical shift database for the various topologies. 13C chemical shifts C8 and C1’ provided an unequivocal way of determining the distribu- G4s are widely observed in guanosine (G-)rich DNA and RNA se- tion, thus leading to reliable and rapid analysis of topology.[1] Ad- quences resulting from the stacking of two/three G-tetrad pseu- ditionally, C1’ CS are found to report on subtle structural features do-planes that are in turn formed by Hoogsteen H-bonds between present within a given topology, aiding in detailed characterization. the G-nucleobases. The relative orientation of the four backbone [1] We further optimized a machine learning based methodology strands defines their classification as parallel, hybrid (3+1), (2+2) using 1H CS to determine structural features of the molecule that antiparallel and antiparallel structures. Additionally, the loops that INTRODUCTION provides an accuracy of ~99% for parallel and ~80% for hybrid (3+1) connect the adjacent backbone strands are also commonly found in SPONSORS topologies.[1] The fidelity of the methodology is demonstrated by its propeller, lateral and diagonal conformations, resulting in range of PROGRAM application to a system that folds into two dissimilar topologies in tertiary structural topologies. Conventional NMR characterization PROGRAM — BY DAY different ionic conditions, providing its first atomic level character- methods are either expensive and/or time-intensive. Our aim was to TOPICS ization.[1] This methodology forms the first step towards creating a develop a CS-based methodology that overcomes these limitations. PRIZE LECTURES database of 13C and 15N chemical shifts for DNA G4s. The insights ob- PLENARY LECTURES Folding of G4s was known to be driven by the formation of back- tained from this study immediately opens up avenues of extending TUTORIAL LECTURE bone grooves, which upon closer inspection unravelled that each the methodology to RNA, map their in vivo structure and monitor INVITED AND PROMOTED LECTURES backbone topology is associated with a specific distribution of the refolding of G4s real-time. POSTERS ADVERTISERS References: [1] RKR. Sannapureddi, MK. Mohanty, AK. Gautam, B. Sathyamoorthy J. Phys. Chem. Lett. 2020, 11(23), 10016-22-365. AUTHOR INDEX Acknowledgments: BS thanks IISER Bhopal and Science and Engineering Research Board (ECR) for the funding. RR thanks IISER Bhopal for funding. MKM thanks DST INSPIRE for funding. PO025 POSTERS / BIOMOLECULES AND INTERACTIONS 178 ALEN ŠADL1, NMR STUDY OF PURINE RICH TANDEM REPEATS MARKO TRAJKOVSKI1, LINKED TO REPLICATION FORK COLLAPSE JANEZ PLAVEC1,2,3 1National Institute of Chemistry, Purine rich tandem repeats form unusual secondary structures linked to replication fork collapse [1]. This phe- Slovenian NMR Centre, Ljubljana, nomenon was demonstrated for different DNA segments, including tandem repeats of the d(CAGAGG) sequence, Slovenia 2University of Ljubljana, Faculty of which three or more tandem repeats are present in more than half of all human chromosomes, some even inside of Chemistry and Chemical important coding sequences like ADAM metallopeptidase, transcription factors and proteins with zinc finger pro- Technology, tein motif. Ljubljana, Slovenia 3EN-FIST Centre of Excellence, Ljubljana, Slovenia We prepared RS4 and SLL1 oligonucleotides comprising three and one repeats of the d(CAGAGG) motif, respective- ly, where extensions at the 5’- and 3’-ends were introduced to promote thermodynamic stability. With an extensive NMR study, we have discovered that RS4 forms structures with stable base-paired stem and AGAG tracts in peculiar conformation, which thus far have not been characterized with high-resolution techniques. We proved that RS4 in the presence of potassium ions folds into a structure that contains two symmetric elements and exhibit two GC and two TA base pairs in Watson-Crick geometry. It is interesting that several 1H NMR signals are extremely broad or could even not be observed, indicating structural equilibrium. Nevertheless, we assigned most of the 1H NMR signals by using isotope-labelled olionucleotides along with constructs specifically designed to carry single-residue substitutions. Unfortunately, for two out of three AGAG segments there is a lack of NOE correlations in NMR spectra of RS4, which is related to extreme broadening of 1H NMR signals. The limited NMR-based insights in structures adopted by RS4, especially the lack of data for the AGAG segments, motivated us to focus on the smaller oligonucle- INTRODUCTION otide SLL1. We found that salt concentration has a great effect on structuring of SLL1, as the higher potassium ion SPONSORS concentrations result in dimer structure formation. With the use of 13C- and 15N-isotopic residue-specific labelling PROGRAM and with the use of 2D NMR methods we assigned most of the 1H NMR signals of SLL1 which allowed further analysis PROGRAM — BY DAY of NOE correlations. While the structural data is consistent with SLL1 adopting a hairpin structure with stable stem, TOPICS details of the AGAG loop remain to be elucidated further in order to explain the evident hydrogen-bonding that PRIZE LECTURES involves imino protons of both guanine residues. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] N. Shastri, Y.C. Tsai, E.J. Brown et al., Mol. Cell. 2018, 72, 222-238. PO026 POSTERS / BIOMOLECULES AND INTERACTIONS 179 PRIMOŽ ŠKET1, PRE-FOLDED STRUCTURES: TJAŠA FRELIH1, FOLDING PATHWAYS OF HUMAN TELOMERIC BAIFAN WANG1 AND JANEZ PLAVEC1,2,3 G-QUADRUPLEXES 1National Institute of Chemistry, Slovenian NMR Guanine-rich (G-rich) DNA regions exhibit potential to form G-quadruplexes in the presence of stabilizing cations, Centre, Ljubljana, Slovenia and demonstrate vast structural diversity and polymorphism. G-rich DNA sequences are present throughout the 2EN-FIST Center of Excellence, human genome including regulatory regions of (onco)genes and telomeric repeats. Consequently, G-quadruplexes Ljubljana, Slovenia 3University of Ljubljana, Faculty are attractive targets for anticancer and antiviral drug development. of Chemistry and Chemical Technology, Ljubljana, Slovenia Understanding the mechanism by which biological macromolecules fold into their functional native conforma- tions represents a problem of fundamental interest. DNA oligonucleotides derived from human telomeric repeat d[TAGGG(TTAGGG) ] and d[TAGGG(TTAGGG) TT] fold into G-quadruplexes through diverse steps. NMR and oth- 3 3 er data acquired at different experimental conditions undoubtedly revealed that both oligonucleotides exhibited defined pre-folded structure(s) already in the absence of cations that are believed to be a starting point of folding process [1]. All determined structures (with base pairs and base triples, with antiparallel chair-like topology as well as hairpin) showed that the first and the second G-tracts are connected in antiparallel orientation. This structural feature could be the main reason for different folding of d[TAGGG(TTAGGG) ] and d[TAGGG(TTAGGG) TT] into 3 3 their final hybrid-1 and hybrid-2 G-quadruplexes, respectively. While formation of hybrid-2 G-quadruplex can proceed directly from antiparallel pre-folded structure, reorientation of the first and the second G-tracts into parallel INTRODUCTION alignment is required for formation of hybrid-1 G-quadruplex. Therefore, immediately after the addition of K+ ions SPONSORS into solution of d[TAGGG(TTAGGG) ], G-quadruplex structure with antiparallel orientation of the first and second 3 PROGRAM G-tracts is likely formed before the final hybrid-1. PROGRAM — BY DAY Studies where we can experimentally characterize structures of pre-folded forms are essential to establish mech- TOPICS anisms of G-quadruplex folding, where pre-folded forms can be on- or off-pathway intermediates and can lead to PRIZE LECTURES development of novel type of selective ligands that will target peculiar structural elements. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] T. Frelih, B. Wang, J. Plavec, P. Sket, Nucleic Acids Res. 2020 48, 2189-2197. AUTHOR INDEX Acknowledgments: This project was supported by Slovenian Research Agency [ARRS, grants P1-0242 and J1-7108]. PO027 POSTERS / BIOMOLECULES AND INTERACTIONS 180 VESNA ŠTIH1,2, NMR STRUCTURAL DETERMINATION OF OxyS sRNA PETER PODBEVŠEK1,3, JANEZ PLAVEC1,2,3 Small non-coding RNAs (sRNAs) are an important class of RNAs in bacteria with regulatory roles in stress response 1 and adaptation to environmental changes [1]. OxyS is a 109 nucleotide long, stable, trans-encoded sRNA found National Institute of Chemistry, Slovenian NMR Centre, Ljubljana, in Escherichia coli [2]. It is regulated by OxyR and is induced in high concentration in response to oxidative stress Slovenia caused by an elevated concentration of hydrogen peroxide (H O ). OxyS is a global regulator affecting the expres-2 2 2 University of Ljubljana, Faculty sion of multiple genes, mainly through direct base-pairing with relevant mRNAs [3]. OxyS is predicted to adopt a of Chemistry and Chemical Technology, Ljubljana, Slovenia secondary structure containing three stem-loops, which was experimentally verified by chemical probing [2]. To 3EN-FIST Centre of Excellence, understand and study base-pairing interactions between target mRNAs and OxyS, more detailed knowledge about Ljubljana, Slovenia its structure is needed. We adopted the divide and conquer approach to separately study the parts of OxyS sequence that are predicted to fold into hairpins. We recorded a series of homonuclear and heteronuclear NMR experiments to determine the solution structures of the isolated stem-loops SL1, SL2 and SL3. Later we in vitro transcribed the full-length OxyS, to study its structure with NMR. We confirmed the presence of stem-loops SL1, SL2 and SL3 in OxyS. Unexpectedly, we identified an additional short hairpin SL4 located in the predicted linker region. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] L. S. Waters, G. Storz, Cell. 2009, 136, 615-628. [2] S. Altuvia et al., Cell. 1997, 90, 43-53. [3] S. Gottesman, Annu. Rev. Microbiol. 2004, 58, 303-331. AUTHOR INDEX Acknowledgments: This work was supported by Janko Jamnik Fund for witty young people in science. PO028 POSTERS / BIOMOLECULES AND INTERACTIONS 181 MARKO TRAJKOVSKI1, STRUCTURAL INSIGHTS INTO NAPHTHALENE DIIMIDE CHIARA PLATELLA2, ANALOGUE INTERACTIONS WITH G-QUADRUPLEXES FILIPPO DORIA3, MAURO FRECCERO3, G-quadruplexes are four-stranded DNA structures, which intriguing 5’- and 3’-end G-quartets of the parallel M2 G-quadruplex. The 1H DANIELA MONTESARCHIO2, biological roles relate to their distinguished features with respect to NMR spectral features in case of NDI-5–M2 G-quadruplex interac- PLAVEC JANEZ1,4,5 the canonical double-stranded helix along with their prevalence tions rendered more detailed investigation with the assignment of 1National Institute of Chemistry, Slovenian in regulatory regions, e.g. promoters of oncogenes and telomeres 1H NMR signals accomplished with the use of site-specifically 13C NMR Centre, Ljubljana, Slovenia 2 [1]. G-quadruplex core comprises stacked G-quartets, each formed and 15N-labelled oligonucleotide synthesis and multi-dimensional University of Naples Federico II, Department of Chemical Sciences, Naples, by four Hoogsteen hydrogen-bonded guanine residues. The loops NMR experiments. The determination of NDI5–M2 G-quadruplex Italy connecting the residues in the core as well as the four stems can be binding stoichiometry afforded the value of 1:3, while at higher ra-3University of Pavia, Department of diverse in terms of length, composition and progression, altogether tios, i.e. at excess of NDI-5, a complex with ‘ligand shell’ features is Chemistry, Pavia, Italy 4EN→FIST Centre of Excellence, Ljubljana, making the G-quadruplexes family extremely polymorphic. Such formed, whereby ligand molecules interact with M2 G-quadruplex Slovenia broad variety of structures adopted by guanine-rich DNA is import- and/or pre-bound ligand. To evaluate the roles of the residues con- 5University of Ljubljana, Faculty of Chemistry and Chemical Technology, ant in context of the evolving therapeutic strategies that rely on dru- necting the guanines in G-quartets, we utilized natural-to-abasic Ljubljana, Slovenia gability of specific G-quadruplex forming regions [2]. residue substitutions at the loop regions that preserved the paral- lel stranded topology of M2 G-quadruplex. The NMR-based assess- Motivated by naphthalene diimide analogue NDI-5 anticancer ac- ment showed that loops modulate the secondary binding of NDI-5, tivity that relates to G-quadruplex binding [3] we explored the un- but are not decisive for the predominant interactions [4]. derlying structural details. We investigated NDI-5 interactions with two G-quadruplex-forming DNA, namely M2 and m-tel24, charac- The present results will help in optimizing properties of G-quadru- INTRODUCTION teristically adopting parallel and hybrid topology, respectively. For plex-binding naphthalene diimide analogues aiming to fulfill the SPONSORS m-tel24 we observed stacking of NDI-5 only at the 5’-end. On the premises of the novel approaches for treatment of different cancers. PROGRAM other hand, NMR results revealed ligand interactions at both, the PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] D. Varshney, J. Spiegel, K. Zyner, D. Tannahill, S. Balasubramanian, Nat. Rev. Mol. Cell. Biol. 2020, 21, 459-474. [2] N. Kosiol, S. Juranek, P. Brossart, A. Heine and K. Paeschke, Mol. POSTERS Cancer. 2021, 20, 40. [3] C. Platella, V. Pirota, D. Musumeci, F. Rizzi, S. Iachettini, P. Zizza, A. Biroccio, M. Freccero, D. Montesarchio, F. Doria, Int. J. Mol. Sci. 2020, 21, 1964. [4] C. Platella, M. ADVERTISERS Trajkovski, F. Doria, M. Freccero, J. Plavec, D. Montesarchio, Nucleic Acids Res. 2020, 48, 12380-12393 AUTHOR INDEX Acknowledgments: The authors acknowledge the CERIC-ERIC consortium for access to NMR facilities. PO029 POSTERS / BIOSOLIDS 182 EKATERINA BURAKOVA, A STATISTICS-DRIVEN NMR APPROACH TO SURESH K. VASA, SITE-SPECIFIC ANALYSIS OF STATIC PROTEIN RASMUS LINSER Faculty of Chemistry and Chemical DISORDER Biology, Technical University of Dortmund, Protein conformational disorder is vital for functionality of a large part of cellular processes and can translate into Dortmund, Germany heterogeneity accessible by solid-state NMR.[1] Understanding the conformational distribution of protein (mis) folding or reaction intermediates on a single-residue level can provide insights useful both from a biological as well as a pharmacological perspective. Disorder in solid-state protein samples has been tackled by various NMR techniques [1]. Here we present two ap- proaches to reconstruction of residue-specific dihedral angle distribution based on analysis of 4D hCBCANH cor- relations. The first approach is based on a comparative analysis of chemical-shift distributions with the relational database PACSY [2]. The second approach involves dihedral angle predictions for the entire 4D peak using TALOS-N [3]. We test both methods on a highly heterogeneous sample of GGAGG pentapeptide. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] A. B. Siemer, Solid State Nucl. Magn. Reson. 2020, 160, 101643. [2] W. Lee et al, J Biomol NMR. 2012, 54,169–179. [3] Y. Shen, A. Bax. J. Biomol. AUTHOR INDEX NMR. 2013, 56, 227–241 PO030 POSTERS / BIOSOLIDS 183 MORGANE CALLON1, PROTON-DETECTED SOLID-STATE NMR ALEXANDER A. MALÄR1, SPECTROSCOPY AT HIGHEST FIELD: SARA PFISTER1, VÁCLAV RÍMAL1, MARCO E. WEBER1, THE GAIN IN RESOLUTION AT 1200 MHZ THOMAS WIEGAND1, Progress in biomolecular NMR spectroscopy is driven by increasing magnetic-field strengths leading to improved JOHANNES ZEHNDER1, resolution and sensitivity of the NMR spectra. We here investigate proton-detected solid-state NMR spectra of a RICCARDO CADALBERT1, variety of biomolecular systems recorded with the recently available superconducting magnet operating at 1200 LAURIANE LECOQ2, MHz proton resonance frequency using magic-angle spinning (MAS) frequencies of 100 kHz. Among them are the ANAHIT TOROSYAN1, MICHAEL deuterated Hepatitis B virus core protein [1], the protonated Rpo4/7 protein complex of two subunits of archaeal NASSAL3, MATTHIAS ERNST1, RNA polymerase II [2] and the protonated amyloid fibrils of the fungal prion HET-s (218-289) [3,4]. ANJA BÖCKMANN2, BEAT H. MEIER1 The spectra reveal a significant gain in resolution compared to spectra recorded at 850 MHz. The linear improve-1 ment of the line width expressed in ppm with increasing B field by a factor of 0.7 (ratio of the two magnetic fields) Physical Chemistry, ETH Zürich, Zürich, 0 Switzerland was shown to be achieved for all the samples. For side-chain protons (in particular CH and CH protons) an even 2 3 2Molecular Microbiology and Structural stronger narrowing was observed as the strong couplings are partially suppressed by the increased chemical-shift Biochemistry,Université de Lyon, Lyon, France 3 separation. This gives access to structure and dynamics studies of aliphatic protons in fully protonated systems. Dept. of Medicine II / Molecular Biology, University of Freiburg, Freiburg, Germany The longer T ’ times, measured for the bulk amide protons at the highest field further allow, in combination with 2 faster MAS, to use J-coupling based sequences to observe dynamic domains of biomolecules that currently escape detection using CP-based magnetization transfer experiments. INTRODUCTION We recorded as well carbon-13 experiments, and show that in 13C spectra substantially higher numbers of peaks can SPONSORS be resolved compared to 850 MHz. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES References: [1] L. Lecoq, et al., Frontiers in Molecular Biosciences. 2019, 6, 58. [2] A. Torosyan et al. , Frontiers in Molecular Biosciences. 2019, 6, 100-108. [3] C. PLENARY LECTURES Wasmer et al. , Science. 2007, 319, 1523-26 [4] H. van Melckebeke et al., JACS. 2010, 132, 13765-75. TUTORIAL LECTURE Acknowledgments: Financial support by the ETH Zurich, the Department of Chemistry and Applied Biosciences, and the ETH foundation have been essential INVITED AND PROMOTED LECTURES for obtaining the spectrometer. The scientific projects featured are supported by an ERC Advanced Grant (B.H.M., grant number 741863, Faster), by the Swiss National Science Foundation (B.H.M., grant number 200020_159707 and 200020_188711), an ETH Research Grant ETH-43 17-2 (T.W.), the Günthard Stiftung POSTERS (M.W.), the French Agence Nationale de Recherches sur le Sida et les hépatites virales (ANRS, ECTZ71388 & ECTZ100488), the CNRS (CNRS-Momentum 2018), ADVERTISERS the LABEX ECOFECT (ANR-11-LABX-0048) within the Université de Lyon program Investissements d’Avenir (ANR-11-IDEX-0007). We thank Dr. Patrick AUTHOR INDEX Wikus and Dr. Rainer Kümmerle from Bruker Schweiz AG for their support. POSTERS / BIOSOLIDS PO031 184 MAIK DERKS1,2, THE MODE OF ACTION OF PLECTASIN & VARIANTS IN SHEHRAZADE JEKHMANE1, MEMBRANES JOÃO MEDEIROS -SILVA1, VICKY CHARITOU1, BAREND ELENBAAS1, Plectasin is an antimicrobial peptide (AMP), belonging to the cysteine stabilized alpha-beta (CSaß) defensins first MAREK PRACHAR1, KAMAL discovered back in 2005 in the fungus Pseudoplectania nigrella. [1] Plectasin and its variants (e.g. NZ2114, MP1102) TEHRANI3, NATHANIEL MARTIN3, have excellent bactericidal activity against a variety of gram positive bacteria. This includes some clinically relevant EDWIN VELDHUIZEN4, strains, among which MRSA, infamous in hospitals for causing difficult to treat infections. Binding of peptidogly-STEPHEN COCHRANE5, can-precursor Lipid II (LII) is thought to be the mechanism of action of these peptides [2] but much remains unclear MARKUS WEINGARTH1, about the structural details. EEFJAN BREUKINK2 1 Unfortunately, studying the atomic structure of peptide-Lipid II complexes in their native environment, a mem- NMR spectroscopy, Utrecht university, The Netherlands brane, by conventional techniques -such as X-ray diffraction- is challenging. Furthermore, without these data the 2Membrane Biochemistry and Biophysics, Utrecht development of these drugs is like stumbling in the dark, hampering the transition to the clinic of these high-po-University, The Netherlands 3 tential candidates. By studying the Plectasin-LII complex in membranes with solid-state NMR we show a mark- Infectious Diseases and Immunology, Utrecht University, The Netherlands edly different binding mode than previously found in micelles. In particular, we find that the aß-loop is critical 4Microbial Technology and Health, Leiden University, for LII binding and forms a rigid ‘clamp’. 13C spin diffusion experiments with universally 13C labeled Plectasin and The Netherlands 5Bioorganic Chemistry, Queen’s University Belfast, LII identify different residues in direct contact with LII. Furthermore, Plectasin-LII complexes aggregate and form United Kingdom micrometer-sized clusters that are clearly visible even with microscopy, potentially pointing to an additional mode of action. Variants of Plectasin with altered microbial scope such as NZ2114 are modified not near the proposed binding site, but around a negatively charged on the opposite side of the peptide. We, therefore, hypothesize that INTRODUCTION these modifications change the aggregation kinetics and thereby influence antimicrobial activity. This is supported SPONSORS by the observation that the NZ2114-LII complex is more dynamic compared to Plectasin. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] P. H. Mygind, R. L. Fisher, H. H. Kristensen, Nature. 2005, 437 (7061), 975-80 [2] T. Schneider, T. Kruse, H. H. Kristensen, Science. 2010, 328 AUTHOR INDEX (5982), 1168-72. PO032 POSTERS / BIOSOLIDS 185 SIMON FRIDOLF, GM3 LIPIDS STIMULATE CO-ASSEMBLY OF VESICLES EMMA SPARR, AND ALPHA-SYNUCLEIN DANIEL TOPGAARD Division of Physical Chemistry, Parkinson’s disease is characterized by co-aggregates of the presynaptic protein alpha-synuclein and lipids in the Lund University, Lund, Sweden brains of patients [1]. This process could cause disease by alpha-synuclein fibril- or oligomer toxicity, depletion of functional alpha-synuclein or by damaging lipid membranes [2]. We used NMR spectroscopy together with cryo-transmission electron microscopy to investigate co-assemblies formed when monomeric alpha-synuclein is aggregating in the presence of POPC vesicles containing ganglioside GM3, a bio-synthetic precursor for brain gangliosides [3]. Our results show that lipids co-assemble with alpha-synuclein to form μm-sized amorphous aggregates, whose properties depend on the lipid-protein ratio in the incubation mixture and the fraction of ganglioside GM3 in vesicles. Solid state 13C MAS NMR analysis of the aggregates together with quantitative solution 1H NMR of superna-tants show that GM3-rich vesicles stimulate more lipids to co-assemble with alpha-synuclein, in a manner that is selective for GM3 over POPC. From measurements of 1H-13C residual dipolar couplings, 31P chemical shift anisotropy, and 13C relaxation rates – reporting on molecular segment reorientation on time-scales from nanoseconds to milliseconds – we deduce that co-assembly with alpha-synuclein induces curvature of the lipid bilayers and slows down reorientation about the molecular long-axis and transitions between conformational isomers. The present findings add insight into the structure and dynamics of lipid/alpha-synuclein co-assemblies and may INTRODUCTION also hint at how ganglioside/alpha-synuclein interactions could have an effect on the structure-function-toxicity SPONSORS relationship related to Parkinson’s disease. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Shahmoradian, S. H. et al., Nature Neurosci. , 2019, 22, 1099-1109. [2] Reynolds, N. P. et al., J. Am. Chem. Sci., 2011, 133, 19366-19375. [3] Schnaar, R. L., Gerardy-Schahn, R., Hildebrandt, H., Physiol. Rev. , 2014, 94, 461-518. INVITED AND PROMOTED LECTURES Acknowledgements: This study was funded by a project grant from the Knut and Alice Wallenberg Foundation (Emma Sparr and Daniel Topgaard). The POSTERS authors thank Göran Carlström (Centre for Analysis and Synthesis, Lund University) and Anna Carnerup (Division of Physical Chemistry, Lund University) ADVERTISERS for technical assistance with solid-state NMR and cryo-TEM experiments, Katja Bernfur (Biochemistry and Structural Biology, Lund University) for providing AUTHOR INDEX purified alpha synuclein and Carl Grey (Division of Biotechnology, Lund University) for mass spectroscopic analysis of the purchased GM3. PO033 POSTERS / BIOSOLIDS 186 ARUN KUMAR INSIGHTS INTO SITE-SPECIFIC DYNAMICS OF RNA BY SREEMANTULA1, SOLID-STATE NMR SPECTROSCOPY JOHN KIRKPATRICK1,2, ALEXANDER MARCHANKA1 The study of biomolecular dynamics provides information about a multitude of cellular processes including folding, 1Biomolekulares Wirkstoffzentrum (BMWZ), binding affinity, allosteric regulation and enzymatic catalysis. Magic-angle spinning solid-state NMR (MAS ssNMR) Leibniz University Hannover, Hannover, is able to characterize motions on the pico- second-to-millisecond timescale and thus gives access to important Germany. 2NMR-based Structural Chemistry, biological dynamic processes ranging from bond fluctuations to folding events [1]. Despite significant progress in Helmholtz Centre of Infection Research, the research of protein structure and dynamics by ssNMR, corresponding studies of RNA remain at a relatively early Braunschweig, Germany stage due to a lack of appropriate experimental protocols and the strong chemical similarity of the four-nucleotide building-blocks, which inevitably leads to spectral crowding [2]. In recent years we have developed the methods that allowed us to obtain the first structures of an RNA and a protein–RNA complex solely by ssNMR [3,4]. Here we report for the first-time site-specific relaxation data for RNA measured by ssNMR spectroscopy on a uniformly 13C,15N labeled sample. We have measured the 15N longitudinal relaxation ( T ) of amino and imino nitrogens of all nu-1 cleotides in the structured regions of a 26mer box C/D RNA in complex with L7Ae protein [3] at different MAS rates. On average, the 15N T values of the amino nitrogens are significantly shorter than those of the imino nitrogens, 1 indicative of faster relaxation of the former due to a stronger overall 15N-1H dipolar interaction. On a nucleotide-specific basis, significantly shorter T values are reported for the non-canonical region of the RNA (loop), whereas those 1 for nucleotides in the canonical elements (stem) and the protein-binding region (kink-turn) are longer, implying reduced flexibility and increased stability in the canonical stem-loop and kink-turn regions. Experimental chal- INTRODUCTION lenges in measuring 15N T relaxation of RNA by ssNMR are discussed and the contribution of spin-diffusion to the 1 SPONSORS relaxation rates is assessed. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] J. R. Lewandowski, H. J. Sass, S. Grzesiek, M. Blackledge, and L. Emsley, J. Am. Chem. Soc. , 2011, 133, 42, 16762–16765. [2] A. K. Sreemantula, A. ADVERTISERS Marchanka, Biochem. Soc. Trans. , 2020, 48, 1077-1087. [3] A. Marchanka, B. Simon, G. Althoff-Ospelt, and T. Carlomagno, Nat. Commun. , 2015, 6, 1–7. [4] M. AUTHOR INDEX Ahmed, A. Marchanka, T. Carlomagno, Angew. Chem. Int. Ed, 2020, 59, 6866-6873. PO034 POSTERS / BIOSOLIDS 187 CHARLOTTE WEISS1, STRUCTURAL STUDY OF WILD TYPE FAT10 BY NICOLA CATONE2, MAS NMR SPECTROSCOPY VENKATA SUBBARAO REDROUTHU1, Human leukocyte antigen (HLA)-F adjacent transcript 10 (FAT10) diffraction and solution NMR of the individual stabilized, cysteine-MARCUS GROETTRUP2,3, is a protein exclusively found in mammals [1]. FAT10 is restricted free domains reveal that both domains fold similarly to ubiquitin, GUINEVERE MATHIES1 to organs and cells of the immune system [2,3]. However, FAT10 but exhibit completely different electrostatic surface potentials to 1Department of Chemistry, expression gets induced in other types of tissue by the synergistic each other as well as to ubiquitin translating into diverse non-co-University of Konstanz, Konstanz, action of tumor necrosis factor alpha (TNF-a) and interferon (IFN)γ valent interactions. As the stabilized version of FAT10 and its con-Germany; 2Biotechnology Institute Thurgau, Kreuzlingen, Switzerland; under inflammatory conditions [4,5,6]. jugates are degraded at slower rates [10], a structural study of wild 3Department of Biology, University of type FAT10 will likely provide new, biologically relevant informa- Konstanz, Konstanz, Germany The protein belongs to the ubiquitin-like modifier (ULM) family tion. MAS NMR spectroscopy is ideally suited for this purpose. which is characterized by a common three-dimensional structure known as the β-grasp fold [7]. FAT10 itself consists of two ubiqui- Since FAT10 has not been studied by MAS NMR before, we have tin-like domains which are connected by a flexible linker [8]. The started our structural studies at the N-domain. We successfully ex-free C-terminal diglycine motif is used for covalent attachment to pressed and purified the uniformly 13C-15N-labelled, cysteine-free substrate proteins. Beside ubiquitin, FAT10 is the only ULM that N-domain of FAT10. A protocol for growing microcrystals was de-targets its substrate proteins for fast and direct degradation by the veloped and resulted in high-resolution 1H-13C CP spectra of the mi-26S proteasome. Mono-FATylation is sufficient for degradation and crocrystalline FAT10 N-domain in natural abundance at 400 MHz. FAT10 is degraded along with its substrates [9]. At 800 MHz, good quality 13C-13C correlation spectra of the uniform- ly 13C-15N-labelled, cysteine-free N-domain of FAT10 were obtained. INTRODUCTION The extremely instable FAT10 tends to precipitate and for this rea- 13C-15N ZF TEDOR spectra at 800 MHz will follow in the near future. SPONSORS son, the cysteine residues were replaced in a structural study of PROGRAM the protein [10]. High-resolution structures determined by X-ray PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] M. Groettrup, et al., Trends in Biochemical Sciences. 2008, 33, 230-237. [2] E. E. M. Bates, et al., European Journal of Immunology. 1997, 27, 2471-2477. [3] C. G. L. Lee, et al. INVITED AND PROMOTED LECTURES Oncogene. 2003, 22, 2592-2603. [4] Y. C. Liu et al., Proc. Natl. Acad. Sci. USA. 1999, 96, 4313-4318. [5] S. Raasi, G. Schmidtke, R. de Giuli, M. Groettrup, European Journal of Immunology. 1999, 29, 4030-4036. [6] S. Lukasiak, et al. Oncogene. 2008, 27, 6068-6074. [7] S. Vijay-Kumar, C. E. Bugg, W. J. Cook, Journal of Molecular Biology. 1987, 194, 531-544. [8] W. Fan, W. Cai, S. Parimoo, POSTERS G. G. Lennon, S. M. Weissman, Immunogenetics. 1996, 44, 97-103. [9] M. S. Hipp, B. Kalveram, S. Raasi, M. Groettrup, G. Schmidtke, Molecular and Cellular Biology. 2005, 25, 3483-3491. ADVERTISERS [10] A. Aichem, S. Anders, N. Catone, P. Roessler, S. Stotz, A. Berg, R. Schwab, S. Scheuermann, J. Bialas, M. C. Schuetz-Stoffregen, G. Schmidtke, C. Peter, M. Groettrup, S. Wiesner, Nature AUTHOR INDEX Communications. 2018, 9, 3321. PO035 POSTERS / BIOSOLIDS 188 ANIKA WURL1, STRUCTURE AND DYNAMICS OF MARIA OTT2, LONG N-ALKANES IN LIPID BILAYERS ALFRED BLUME3, TIAGO M FERREIRA1 DETERMINED BY SOLID-STATE NMR 1Institute of Physics, 2Institute of Biochemistry and Biotechnology, Understanding the fate of hydrophobic polymers in cells is important due to increasing environmental concerns, 3Institute of Chemistry, Martin-Luther such as the accumulation of nanoplastics in our oceans. Surprisingly, little is known about the potential incorpora-University Halle-Wittenberg, Halle, tion of long hydrophobic chains into the hydrophobic core of cellular membranes [1]. Germany Due to their simplicity, n-alkanes are thought a convenient starting point for investigating combined systems of lipids and hydrophobic polymers. However, contrary to shorter n-alkanes, the mixing of long alkanes (20 carbons and longer) with lipid bilayers has not been studied in detail [2,3]. Solid-state NMR is well suited to measuring the dynamics of lipid bilayers and incorporated molecules. We are using 1H-13C dipolar recoupling solid-state NMR [4] and static 2H/31P NMR, combined with x-ray scattering and MD simulations to characterize the structure and dynamics of various n-alkane/phospholipid membranes, and to determine how hydrophobic chain length (up to 30 carbons) and hydration affect the membrane molecular properties. The combination of experiments and simulations used enables to describe the systems with unprecedented atomistic detail. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] D. Bochicchio, E. Panizon, L. Monticelli, G. Rossi, Sci Rep. 2017, 7, 6357-6365. [2] J.M. Pope, L.A. Littlemore, P.W. Westerman, Biochim. Biophys. ADVERTISERS Acta. 1989, 980, 69-76. [3] H. Usuda, M. Hishida, E.G. Kelley, Y. Yamamura, M. Nagao, K. Saito, Phys. Chem. Chem. Phys. 2020, 22, 5418-5426. [4] S.V. Dvinskikh, AUTHOR INDEX V. Castro, D. Sandström, Phys. Chem. Chem. Phys. 2005, 7, 607-613. PO036 POSTERS / BIOMOLECULAR DYNAMICS 189 KALYAN S. CHAKRABARTI1, SIMON OLSSON2, SUPRIYA A LITMUS TEST USING HIGH-POWER PRATIHAR3, KARIN GILLER3, KERSTIN OVERKAMP3, KO ON RELAXATION DISPERSION FOR LEE4, VYTAUTAS GAPSYS5, KYOUNG-SEOK RYU4, BERT L. DE GROOT5, FRANK NÓE6,7,8, STEFAN BECKER3, DONGHAN LEE9, CLASSIFICATION OF RECOGNITION THOMAS R.WEIKL10, CHRISTIAN GRIESINGER3 MECHANISMS OF TRANSIENTLY BINDING 1Division of Sciences, Krea University, India. 2Department of Computer Science and Engineering, Chalmers University of Technology, Gothenburg, Sweden. PROTEINS 3Department of NMR Based Structural Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany. 4 Molecular recognition in proteins is critical for all biological functions, and yet, delineating Research Center for Bioconvergence Analysis, Korea Basic Science Institute, Ochang-Eup, Cheongju-Si, Chungcheongbuk-Do, South Korea. its mechanism is challenging, especially when recognition happens within microseconds 5Department of Theoretical and Computational Biophysics, Max Planck Institute for [1]. We present a novel theoretical and experimental framework to distinguish between two- Biophysical Chemistry, Göttingen, Germany. 6Department of Mathematics and Computer Science, Freie Universität Berlin, Berlin,Germany. state vs three-state binding, including conformational selection and induced fit [2], based 7Department of Physics, Freie Universität Berlin, Berlin, Germany. on straightforward kinetic experiments using NMR high-power relaxation dispersion [3], 8Department of Chemistry, Rice University, Houston, TX, USA. sensitive to single-digit microseconds. The novel framework predicts that conformational 9Department of Medicine, James Graham Brown Cancer Center, University of Louisville, Louisville, KY,USA. selection prevails on ubiquitin’s paradigmatic interaction with the SH3c domain from an 10Department of Theory and Bio-Systems, Max Planck Institute of Colloids and Interfaces, adapter protein [4]. We then reveal the residues that engage in the conformational selection Potsdam, Germany; mechanism using molecular dynamics simulations and Markov state modeling [5, 6]. The kalyan.chakrabarti@krea.edu.in novel framework is robust and expandable for implementation in other binding scenarios with the potential to show that conformational selection might be the design principle of the hubs of interaction networks. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] K. S. Chakrabarti, et al. , Cell Rep. 2016, 14, 32. [2] F. Paul, T. R. Weikl, PLoS Comput. Biol. 2016, 12, e1005067. [3] INVITED AND PROMOTED LECTURES J. G. Reddy, et al. , J. Biomol. NMR 2017, 70, 1. [4] O. F. Lange, et al. , Science 2008, 320, 1471. [5] J.-H. Prinz, et al. , J. Chem. Phys. POSTERS 2011, 134, 174105. [6] C. Schütte, A. Fischer, W. Huisinga, P. Deuflhard, J. Comp. Phys. 1999, 151,146. ADVERTISERS Acknowledgements: Intramural Research Fellowship, Krea Univerisity, Max Planck Society, Max Planck Institute for AUTHOR INDEX Biophysical Chemistry. PO037 POSTERS / BIOMOLECULAR DYNAMICS 190 NITIN KACHARIYA1,2, STRUCTURAL ANALYSIS OF 3’ SPLICE SITE RALF STEHLE1,2, RNA RECOGNITION BY U2AF2-SF1 COMPLEX IN MICHAEL SATTLER1,2 1Bayerisches NMR Zentrum, SOLUTION Department Chemie, Technische Universität München, Germany Recognition of non-coding introns from the pre-mRNA by splicing assembly is a crucial step of gene regulation in 2Institute of Structural Biology, eukaryotes. Several multi-domain complex proteins participate in intron recognition at the early stage of complex Helmholtz Zentrum München, Neuherberg, Germany E. In this assembly, 3’ splice site (SS) of the intron is recognized by the U2 auxilliary factor heterodimer (U2AF), comprised of a small (35kDa, U2AF1), and large (65 kDa, U2AF2) subunit and splicing factor 1 (SF1), which bind to the conserved yAG splice site (3’ SS), the poly pyrimidine tract (PY-tract), and the branch point site (BPS), respectively. Multiple protein-protein interactions also stabilize the assembly. For example, the U2AF1 UHM (U2AF homology motif) domain binds to the U2AF2 ULM (UHM ligand motif), while the U2AF2 UHM domain binds to the SF1 ULM motif1,3. Additionally, SF1 has a helix-helix domain preceding the KH-QUA2 RNA binding region with a conserved RSPSP motif, which undergoes post-translational modification2. Altogether, the multi-domain complex assembly of U2AF2-SF1 is around 120kDa in size and the structures of individual domains are solved by NMR and X-ray crystallography. However, the overall architecture of the U2AF2-SF1-RNA complex structure is not known. Here, we characterize the architecture of the SF1-U2AF2-RNA complex by combining biochemical and NMR ex- periments with SAXS data. We studied binding of a range of RNA motifs to the U2AF2-SF1 complex representing variable strength of cis regulatory RNA motifs. Depending on the RNA binding affinities the overall architecture of INTRODUCTION the U2AF2-SF1 RNA complex adopts compact, semi-compact to open conformation in solution. We use the para- SPONSORS magnetic relaxation enhancement (PRE), pseudo-contact shifts (PCS) and residual dipolar coupling (RDC) data to PROGRAM determine inter-domain distances and orientation restraints, which are used for rigid body modeling to calculate PROGRAM — BY DAY the ensembles of U2F2-SF1-RNA ternary complex. Analysis of 31P NMR spectra of phosphorylated SF1 shows differ- TOPICS ential line broadening of two 31P signals upon complex formation with U2AF2 and RNA, suggesting a long range PRIZE LECTURES effects, that are mirrored by corresponding NMR chemical shift perturbations(CSPs) in U2AF2. In summary, our PLENARY LECTURES results provide the structural insight of RNA recognition by U2AF2-SF1 complex and effect of phosphorylation. TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Zhang Y. et al., Nucleic Acid Research 2013, 41:2, 1343–1354. [2] Lipp, J. et al., Nat Struct Mol Biol 2015, 22, 611–617. [3] Chatrikhi R. et al., Biophysical AUTHOR INDEX Journal 2016 , 111:12, 2570-2586. PO038 POSTERS / BIOMOLECULAR DYNAMICS 191 ATUL RANGADURAI1, EXTENDING THE SENSITIVITY OF CEST NMR HONGLUE SHI2, SPECTROSCOPY TO MICRO-TO-MILLISECOND HASHIM M. AL-HASHIMI1,2 1Department of Biochemistry, Duke DYNAMICS IN NUCLEIC ACIDS USING HIGH-POWER University School of Medicine, Durham, NC, USA RADIO-FREQUENCY FIELDS 2Department of Chemistry, Duke University, Durham, NC, USA Biomolecules undergo motions on the micro-to-millisecond timescale to adopt low-populated transient states that play important roles in folding, recognition, and catalysis. NMR techniques, such as Carr–Purcell–Meiboom–Gill (CPMG), chemical exchange saturation transfer (CEST), and R are the most commonly used methods for character-1r izing such transitions at atomic resolution under solution conditions. CPMG and CEST are most effective at characterizing motions on the millisecond timescale. While some implementations of the R experiment are more broad-1r ly sensitive to motions on the micro-to-millisecond timescale, they entail the use of selective irradiation schemes and inefficient 1D data acquisition methods. Herein, we show that high-power radio-frequency fields can be used in CEST experiments to extend the sensitivity to faster motions on the micro-to-millisecond timescale. Given the ease of implementing high-power fields in CEST, this should make it easier to characterize micro-to-millisecond dynamics in biomolecules. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PO039 POSTERS / BIOMOLECULAR DYNAMICS 192 RAJESH KUMAR REDDY DOES DUPLEX IMPACT QUADRUPLEX: SANNAPUREDDI1, CHARACTERIZATION USING NMR AND MD MANISH KUMAR MOHANTY1, LOIC SALMON2, SIMULATIONS BHARATHWAJ SATHYAMOOTHY1 In genome, nucleic acids form duplex and non-duplex structures, where they coexist. G-quadruplexes (G4s) are well 1 studied non-duplex structures formed by guanine rich sequences. They play pivotal role in genome stability, telo-Department of Chemistry, Indian Institute of Science Education and Research Bhopal, mere maintenance, transcription regulation, protein chaperoning and liquid-liquid phase separation.[1] G4s are India. considered to be potential drug target for cancer and viral infections.[2] So far G4s were studied individually, struc-2Very High Field NMR Center, CNRS, ENSL, tural characteristics of G4s in presence of duplex has not been well characterized. We designed quadruplex-du- UCBL, Lyon University, France plex (QD) systems to mimic QDs formed at promoters and telomeres, where duplex exist on 5’/3’ end G4 forming sequences. 3-plane parallel G4 and dodecamer duplex were attached with varying linker lengths. 5’QD (or 3’QD) represents duplex at 5’-end (or 3’-end) of G4 sequence. Here we present atomistic characterization G4 and impact of adjacent duplex using NMR and molecular dynamic (MD) simulations. Structural and dynamic analysis of G4 pointed out increased flexibility of 5’-tetrad guanines deoxyribose sugars. Interestingly same 5’-tetrad residues form most planar rigid quartet. Trends indicate delicate 3’-tetrad, shows least planar structure. QD systems showed similar trends, yet with chemical shift perturbations (CSP) and varied dynamics. 3’QD show minimal CSP and increasing linker length further decreased CSP. Such changes were rationalized with stacking interactions between tetrads and flanking residues. We observed decreased flexibility of junction INTRODUCTION residues, coaxial stacking between G4 and duplex. Further insights on ligand binding ability to QD systems are SPONSORS provided. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] N. D. Varshney, J. Spiegel, K. Zyner, D. Tannahill, S. Balasubramanian, Nat. Rev. Mol. Cell Biol., 2020 , 21, 459 [2] N. Kosiol, S.Juranek, P. Brossart, AUTHOR INDEX A. Heine, K. Paeschke, Mol. Cancer, 2021, 20(1), 40. PO040 POSTERS / INTEGRATED STRUCTURAL BIOLOGY 193 YING WANG1, SUBSTRATE RECRUITMENT OF THE HEXAMERIC JOHN KIRKPATRICK1,2, MecA-ClpC COMPLEX TERESA CARLOMAGNO1,2 1Centre of Biomolecular Drug Research Regulated proteolysis by ATP-dependent AAA+ proteins is a critical ComK at a low-level. However, when cell-density is high, the degra- (BMWZ), Leibniz University Hannover, component of the protein quality-control system that has evolved in dation of the ComK is blocked by synthesis of an anti-adaptor pro- Hannover, Germany; 2Group of NMR-based Structural all domains of life to maintain cellular homeostasis and to respond tein, ComS, which competes with ComK for binding to MecA and Chemistry, Helmholtz Centre for to external stimuli and environmental changes. In bacteria, the un- allows development of competence by rescuing ComK from degra- Infection Research, Braunschweig, foldase ClpC, which is a member of the Hsp100/Clp family of AAA+ dation [2]. Germany proteins, interacts with its adaptor protein MecA to form a hexam- Despite a number of mechanistic studies on Hsp100/Clp proteins, eric molecular machine that is responsible for controlled unfolding we lack a clear picture of the structural changes occurring at the of protein substrates. Hexameric ClpC can further associate with various stages of the unfolding cycle. For example, the available the protease ClpP to form a complete protein degradation complex crystal structure of the MecA-ClpC complex [3] lacks all the mobile (ClpCP); proteins unfolded by ClpC are translocated to ClpP for pro-elements involved in ATP-binding, hydrolysis and substrate recog- teolytic degradation [1]. nition, leaving open critical questions regarding the mechanisms of As a specific adaptor protein of ClpC, the two-domain protein MecA substrate selection, recognition and unfolding and the coupling of performs a dual role in the unfolding process. It binds substrate these steps to the ATPase activity. proteins by binding to them via its N-terminal domain (NTD), while Here, we report for the first time the in-vitro assembly of the ComK– its C-terminal domain (CTD) interacts with ClpC, promoting its as- MecA–ClpC ternary complex for biochemical and structural analy- sembly into the functional hexameric state. ClpC itself is comprised INTRODUCTION ses. As a step towards obtaining additional high-resolution struc- of two nucleotide-binding domains, D1 and D2. The D1 domain in- SPONSORS tural information, we have performed crystallization of the ternary teracts with MecA, while the D2 domain interacts with ClpP in the PROGRAM complex with a ΔD2 variant of ClpC. In addition, we have undertak- ClpCP complex. PROGRAM — BY DAY en preliminary investigations of both the MecA–ClpC and ComK– TOPICS One of the most important substrates for the ClpCP system is the MecA–ClpC complexes by solution NMR, acquiring methyl-TROSY PRIZE LECTURES transcription factor ComK, which is responsible for competence de- spectra of the MecA component in these ~650kDa and ~700kDa PLENARY LECTURES velopment (uptake of exogenous DNA). Under favourable growth species, respectively. These spectra represent a starting point for TUTORIAL LECTURE conditions, the degradation of ComK via MecA-mediated targeting further NMR-based structural and functional studies of this import- INVITED AND PROMOTED LECTURES to the ClpCP system acts to maintain the cellular concentration of ant system. POSTERS ADVERTISERS AUTHOR INDEX References: [1] J. Liu et al., J. Biol. Chem. 2013, 288, 17597–17608. [2] A. Battesti, and S. Gottesman, Curr Opin Microbiol 2013, 16, 140–147. [3] F. Wang et al. , Nature 2011, 471, 331–337. PO041 POSTERS / IN-CELL MAGNETIC RESONANCE 194 SOPHIE KETTER1, IN SITU LABELING AND DISTANCE MEASUREMENTS OF A. GOPINATH1, MEMBRANE PROTEINS IN E. COLI USING FINLAND AND OX063 O. ROGOZHNIKOVA2, D. TRUKHIN2, TRITYL LABELS V. M. TORMYSHEV2, E. G. BAGRYANSKAYA2, AND In situ structural and dynamical investigations are essential as the memory time (T ), and high stability under reducing environments M B. JOSEPH1 cellular environment can critically impact the conformation and are very favorable for in situ applications. On the other side, their hy-1 function of a protein. Yet such studies represent a challenging task. drophobicity may lead to poor labeling, reduced T , or aggregation. Institute of Biophysics, Department of M Physics and Centre for Biomolecular Electron spin resonance (ESR) spectroscopy combined with site-di- Replacing the FTAM core with the OX063 core increased the water Magnetic Resonance, Goethe University rected spin labeling (SDSL) is emerging as a powerful technique for solubility. Here we spin labeled BtuB with a FTAM and OX063 based Frankfurt, Germany 2 this purpose. labels, all of which bind to the protein with varying linkers through N. N. Vorozhtsov Novosibirsk Institute of Organic Chemistry, SB RAS, Russia a methanethiosulfonate group. We show that both the Finland and Over the past years, we demonstrated a pulsed electron-electron OX063 based trityl labels possess an increased redox stability and double resonance (PELDOR or DEER)-based approach to observe enable specific labeling of BtuB in E. coli with high efficiency. The the structure and conformational changes of outer membrane OX063 labels revealed a long phase memory time close to 5 µs (vs. proteins (OMPs) in intact E. coli and the isolated native outer mem- ~3 µs for the FTAM label) at 100 K when attached to the protein. All brane (OM) [1]. The cobalamin transporter BtuB was labeled with trityl labels enabled in-cell distance measurements between BtuB MTSL and distance measurements were performed to a spin labeled and an orthogonally nitroxide labeled substrate (TEMPO-CNCbl) cobalamin in E. coli and isolated native outer membranes. We also with high selectivity and sensitivity down to a few micromolar of reported distance measurements on doubly labeled BtuB to observe concentration. Overall, all trityls gave relatively narrow distance INTRODUCTION the ligand-induced conformational changes in E. coli [2]. The ni- distributions and did not interfere with ligand binding. The orthog- SPONSORS troxide label (MTSL) suffers from a short lifetime in the reducing onal labeling and PELDOR we demonstrated here would facilitate PROGRAM cellular environment and a high background labeling diminishing the investigation of conformational dynamics and or inter-subunit PROGRAM — BY DAY the overall sensitivity. interactions of key membrane protein complexes in their native en- TOPICS The Finland trityl (FTAM) based labels have emerged as promising vironments [3]. PRIZE LECTURES tags for in situ ESR spectroscopy. Their narrow spectrum, long phase PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] B. Joseph et. al, Nat Protoc. 2019, 14 (8), 2344-2369. [2] B. Joseph et. al, J Am Chem Soc. 2016, 138 (6), 1844-1847. [3] S. Ketter et. al, Chem. Eur. J.. 2021, 27 (7), 2299-2304. PO042 POSTERS / IN-CELL MAGNETIC RESONANCE 195 DANIEL KRAFČÍK1,*, PROBING THE G-QUADRUPLEX-LIGAND EVA IŠTVÁNKOVÁ1, INTERACTIONS IN THE INTRACELLULAR SPACE BY IN- ŠIMON DŽATKO1, PAVLÍNA VÍŠKOVÁ1, CELL NMR SPECTROSCOPY SILVIE FOLDYNOVÁ- TRANTÍRKOVÁ1,2, The 1H-detected in-cell NMR spectroscopy is a powerful technique used to characterize the structural behavior of LUKÁŠ TRANTÍREK1 nucleic acids in living cells [1]. The applications of in-cell NMR studies in human cells have involved double-strand-1 ed and i-motif DNA and hairpin and aptamer RNA. Central European Institute of Technology, Masaryk University, Brno, This study explores the potential of applying 1H and 19F-detected in-cell NMR spectroscopy to profile targeting DNA Czech Republic 2 Institute of Biophysics of the AS CR, G-quadruplexes by ligands. We show that the extension of the original in-cell NMR approach to polymorphic G-qua-v.v.i., Brno, Czech Republic druplex-based targets is not straightforward. The severe signal broadening and overlap in 1H in-cell NMR spectra of * krafcik.daniel@gmail.com polymorphic G-quadruplexes and their complexes complicate their quantitative interpretation. Nevertheless, the 1H in-cell NMR can be used to assess the capability of ligand to bind G-quadruplexes in the native environment. In addition, we tested a recently developed 3,5-bis(trifluoromethyl)phenyl probe to monitor the intracellular G-quadruplex-ligand interactions via 19F-detected in-cell NMR [2]. The 19F-signal from the probe allows to discriminate different G-quadruplex topologies and reveals their number and relative populations. Notably, it also reveals information on G-quadruplex-ligand interaction both in vitro and in cellulo. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] Krafcikova, M.; Dzatko, S.; Caron, C.; Granzhan, A.; Fiala, R.; Loja, T.; Teulade-Fichou, M.-P.; Fessl, T.; Hänsel-Hertsch, R.; Mergny, J.-L.; et al. TUTORIAL LECTURE J. Am. Chem. Soc. 2019, 141, 13281–13285. [2] Bao, H.-L.; Ishizuka, T.; Sakamoto, T.; Fujimoto, K.; Uechi, T.; Kenmochi, N.; Xu, Y. Nucleic Acids Res. 2017, 45, 5501–5511 INVITED AND PROMOTED LECTURES Acknowledgements: This research was funded by the Ministry of Health of the Czech Republic, grant number NV-19-08-00450. The SFT was funded by POSTERS the project SYMBIT (reg. number: CZ.02.1.01/0.0/0.0/15_003/0000477) financed by the European Regional Development Fund and co-financed by the ADVERTISERS Ministry of Education, Youth and Sports of the Czech Republic (MEYS). Access to the research infrastructure was supported by the grant from the MEYS AUTHOR INDEX (CIISB LM2018127). PO043 POSTERS / IN-CELL MAGNETIC RESONANCE 196 SARKA POSPISILOVA1, A THERMO-SENSITIVE GEL MATRIX FOR CELL MICHAELA KRAFCIKOVA1,2, ENCAPSULATION IN BIOREACTORS MATEJ DZUROV3, LUCY VOJTOVA3, Studying biomolecules directly in living cells using the in-cell NMR approach raises the issue of keeping the cells LUKAS TRANTIREK1, at favorable culture conditions inside the NMR cuvette during the experiment. Recently, various types of bioreac-JAN RYNES1* tors have been employed to provide continuous nutrient supply to the sample. To ensure the even distribution of 1Central European Institute of Technology, the fresh cultivation media, cells in the cuvette are usually encapsulated in a porous matrix. Commonly used gels, Masaryk University, Brno, Czech Republic such as alginate or agarose, don’t allow gentle recovery of intact single cells for further assays, e.g., flow cytometry (*rynes@ceitec.muni.cz) 2Institute of Biophysics of the AS CR, v.v.i., analysis. Here, we show an alternative type of matrix with a temperature-sensitive sol-gel transition: liquid at low Brno, Czech Republic temperatures and gel when heated. It is non-toxic to the cells, suitable for in-cell NMR experiments using a bioreac-3Central European Institute of Technology, tor, and allows recovery of single cells without impairing their viability. Brno University of Technology, Brno, Czech Republic INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES Acknowledgments: This work was supported by the interdisciplinary project CEITEC Bridge Fund 2020-2021 and by a grant from the Ministry of Health of POSTERS the Czech Republic (NV19-08-00450) JR and MK were supported by a grant from the Czech Science Foundation (19-26041X) and project SYMBIT (CZ.02.1.01/ ADVERTISERS 0.0/0.0/15_003/0000477) funded by the European Regional Development Fund and Ministry of Education, Youth, and Sports of the Czech Republic (MEYS), AUTHOR INDEX respectively. MEYS is also acknowledged for its support of access to research infrastructure (CEITEC 2020 LQ1601; CIISB-LM2015043). PO044 POSTERS / IN-CELL MAGNETIC RESONANCE 197 ADIL SAFEER1, SOLID-STATE NMR STUDIES OF ARTIFICIAL DAVID BERIASHVILI1, METALLOENZYMES IN WHOLE E. COLI CELLS GERARD ROELFES2, ANDREI GURINOV1, NMR has become a powerful spectroscopic tool to elucidate enzyme structure and dynamics in solution and sol- MARC BALDUS1 id-state NMR. Thus far, such studies have focused on characterizing proteins under in vitro conditions. 1NMR Spectroscopy group, Bijvoet Center for Biomolecular Research, Recently, we could show that in-cell solid-state NMR (see, e.g., Ref. [1]) can be used to probe an artificial metalloen-Utrecht University Utrecht, The zyme (ArM) at atomic level in whole Escherichia coli cells by employing magic-angle spinning (MAS) solid-state (ss) Netherlands 2 NMR spectroscopy. Our studies focused on an ArM comprised of copper(II) phenanthroline bound to the homod- Stratingh Institute for Chemistry, University of Groningen, Groningen, imer of lactococcal multidrug resistance regulator (LmrR) that is capable of catalyzing Friedel-Crafts alkylation of The Netherlands. indoles with high yields and exceptional enantioselectivity[2]. So far, our in-cell ssNMR studies were restricted to using dynamic nuclear polarization (DNP) MAS-ssNMR at com- parably low magnetic field (400 MHz/263 GHz) due to the unfavorable magnetic field-dependence of bi-nitroxide radicals [3]. Here we present experimental results of LmrR expressing cells obtained at 800 MHz/527 GHz DNP conditions, making use of recent advancements in DNP radical design (see, e.g., Ref. [4]) and compare our spectroscopic results to our earlier work [2]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] S. Narasimhan, C. Pinto, A. L. Paioni, J. van der Zwan, G. E. Folkers, M. Baldus, Nat Protoc. 2021, 16, 893-918. [2] S. Chordia, S. Narasimhan, INVITED AND PROMOTED LECTURES A. Lucini Paioni, M. Baldus, G. Roelfes, Angew Chem Int Ed. 2021, 60, 5913-5920. [3] D. Mance, P. Gast, M. Huber, M. Baldus, K. L. Ivanov, J Chem Phys. 2015, POSTERS 142. [4] W. Zhai, A. Lucini Paioni, X. Cai, S. Narasimhan, J. Medeiros-Silva, W. Zhang, A. Rockenbauer, M. Weingarth, Y. Song, M. Baldus, Y. Liu, J Phys Chem B. ADVERTISERS 2020, 124, 9047-9060 AUTHOR INDEX Acknowledgements: Bart Brouwer, Dr. Siddarth Narasimhan. PO045 POSTERS / IN-CELL MAGNETIC RESONANCE 198 PAVLÍNA VÍŠKOVÁ1, CELL CYCLE RESOLVED NUCLEIC ACID IN-CELL NMR EVA IŠTVÁNKOVÁ1, SPECTROSCOPY MICHAELA KRAFČÍKOVÁ2, ŠIMON DŽATKO1, In-cell NMR spectroscopy is a powerful tool for investigating the behavior of nucleic acids inside living cells in a TOMÁŠ LOJA1, close-to-physiological environment [1]. However, this method has only been limited to studies of nucleic acid struc-JAN RYNEŠ1, ture and interactions in the asynchronous cell suspensions. Studying the processes and parameters that are chang-SILVIE FOLDYNOVÁ- ing in the course of cell cycle progression has become essential for understanding the regulation of many cellular TRANTÍRKOVÁ2, functions and human pathologies, including cancer [2,3]. LUKÁŠ TRANTÍREK1 1 This study focused on adapting the in-cell NMR approach for monitoring nucleic acid structure and interaction in Central European Institute of Technology, Masaryk University, Brno, separate cell cycle phases. Using a model example of a double-stranded DNA, we show that the in-cell NMR spec- Czech Republic, troscopy method can be applied to evaluate the nucleic acid behavior in cells synchronized in mitosis and early S 2Institute of Biophysics of the AS CR, phase. v.v.i., Brno, Czech Republic, pavlina.viskova@ceitec.muni.cz INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Dzatko, S. et al., Angew Chem Int Ed Engl 2018, 57 (8), 2165–2169. [2] Alberts, B. et al., Molecular Biology of the Cell. 4th edition 2002. [3] Vermeulen, K. et al., Cell Proliferation 2003, 36 (3), 131–49. INVITED AND PROMOTED LECTURES Acknowledgments: This research was supported by the grant from Czech Science Foundation (19-26041X). MK and SFT were supproted by the project POSTERS SYMBIT (CZ.02.1.01/0.0/0.0/15_003/0000477) funded by the European Regional Development Fund and Ministry of Education, Youth, and Sports (MEYS) ADVERTISERS of the Czech Republic. MEYS is also acknowledged for its support of access to research infrastructure (CIISB-LM2018127; Czech-BioImaging LM2018129; AUTHOR INDEX EATRIS-CZ LM2018133). PO046 POSTERS / SMALL MOLECULES 199 DIMITRIS ARGYROPOULOS EVALUATION OF THE BENEFIT AND INFORMING CAPABILITY AND OF 2D NMR EXPERIMENTS FOR STRUCTURE ELUCIDATION MIKHAIL ELYASHBERG Advanced Chemistry Development (ACD/ USING CASE SOFTWARE Labs) Inc., Toronto, Canada Computer Assisted Structure Elucidation (CASE) has been around benefits and potential problems that are encountered when using for more than 50 years [1,2]. It has experienced a significant boost modern experiments like LR-HSQMBC, in addition to the tradi-after the introduction of routine 2D NMR experiments in the 1990’s, tional HSQC and HMBC. Here, we will also study the influence of as the increased information content offered allowed much more manually adjusting atom properties (such as hybridization and con-complex problems to be addressed. The evolution of computers and nection to heteroatoms). The final example serves for considering the processing power of modern CPUs allowed CASE to be a tool of the inter-relation between HMBC and INADEQUATE when a CASE reference for resolving the unique and unprecedented structures of system is used. natural products[3]. Some common questions encountered when We see that even though the first two structures can be solved with spectroscopists are introduced to CASE are regarding the minimum only the most basic NMR experiments, addition of specialised cor- set of experiments required to solve a problem by CASE and what is relation experiments can decrease the elucidation time dramati-the value and informing capability of advanced NMR experiments. cally. In the third example we see that even though INADEQUATE Although the answers vary depending on the problem complexity, can speed up the elucidation time, this is not of too much benefit; in this poster we will try to tackle both questions, by demonstrating the problem can be solved with only HMBC data in a reasonable a few cases. time, much less than the time required to setup and record the INTRODUCTION We will look at three example structures to answer these questions INADEQUATE. SPONSORS more efficiently: 2-Ethylindanone, Spirodactylone [4] and a xan- Exact details of the type of spectra used, and the calculations per- PROGRAM thone-class natural product [5-6]. In the first example, which is a formed will be shown, as well as comparative tables on the achieved PROGRAM — BY DAY very simple molecule, we will examine the benefits of basic NMR improvements in performance. TOPICS experiments and how they affect the calculation time. The second PRIZE LECTURES example is a challenging natural product, and we will examine the PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] M. Elyashberg, D. Argyropoulos, eMagRes, 2019, 8, 239–254. [2] M. Elyashberg, D. Argyropoulos, Mag. Reson. Chem., doi:10.1002/mrc.5115. [3] M.E. Elyashberg, A.J. Williams. ADVERTISERS “Computer-based Structure Elucidation from Spectral Data. The Art of Solving Problems”, Springer, Heidelberg, 2015, 454. [4] U. Kang, D. Caldwell et al., Org. Lett. 2019, 21, 4750−4753. [5] L. AUTHOR INDEX Boudesocque-Delaye, D. Agostinho et al., J. Nat. Prod. , 2015, 78 (4), 597–603. [6] J. J. Omolo, V. Maharaj et al., J. Nat. Prod. 2012, 75, 1712−1716. PO047 POSTERS / SMALL MOLECULES 200 MARGHERITA BAZZONI1§, NMR ELUCIDATION OF THE DYNAMIC FORMATION ANDREA SECCHI1, AND REORIENTATION OF FLUORESCENT CALIX[6] ARTURO ARDUINI1 1Dipartimento SCVSA, Università di ARENE BASED ISOMERIC PSEUDOROTAXANES Parma Parco Area delle Scienze 17/A, 43124, Parma, Italy The stimuli-triggered dynamic equilibrium of supramolecular complexes has a crucial role in the growing interest §Present address: CEISAM, CNRS for supramolecular chemistry and is the base for most potential applications. In the case of the encapsulation of a UMR6230, Université de Nantes, 44300 Nantes, France fluorescent dye, the displacement of the relative complexation equilibrium can lead to interesting optical properties. For example, the optical properties of the dye can be tuned as a function of its geometrical arrangement inside the host molecule [1], and the dye spatial confinement in the complex may prevents/reduces quenching phenomena [2]. However, the elucidation of dynamic equilibria is not often trivial. Mixing data obtained from multiple analytical techniques is needed, and among these NMR experiments are crucial for structural elucidation, even though these systems often lead to complicated spectra with significant overlap and broad signals. Here we exploit a thorough NMR characterization to investigate the encapsulation of a stilbazionium salt into a non-palindromic tris-(N)phenylureido calix[6]arene [3]. The mixing of the two components in a low polar solvent leads to the formation of two isomeric complexes endowed with different optical properties and in a temperature triggered dynamic equilibrium between each other. The NMR spectra taken on a 1:1 host/guest mixture were all characterized by complex signals patterns due both to a high conformational mobility of the complexed species and to exchange processes occurring on the NMR time scale. A plethora of NMR experiments, including DOSY, RO- INTRODUCTION ESY and VT experiments, were exploited to overcome the difficulties related to peak overlap and broad lineshapes. SPONSORS These experiments made it possible to assess that the two complexes differ for the orientation of the guest inside PROGRAM the host, the DOWN isomer shows the diethylamino group in proximity of the lower rim of the host, while in the PROGRAM — BY DAY UP isomer this group is oriented at the upper rim of the host. The spectroscopic optical measures on the mixture TOPICS allowed to assess the two spectroscopic behaviors present in solution but are blind on their assignment to a specific PRIZE LECTURES geometry. A kinetic follow up of the temperature triggered equilibration process performed through both UV-vis PLENARY LECTURES and fluorescence and NMR experiments allowed to assess which optical properties belong to which isomer. TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] N. Mourtzis, K. Eliadou, K. Yannakopoulou, Supramol. Chem. 2004,16, 587–593; [2] R. N. Dsouza, U. Pischel, W. M. Nau, in Supramolecular ADVERTISERS Photo-chemistry, Ramamurthy, V., Inoue, Y., Eds.; John Wiley & Sons, Inc.: 2011, 87–114; [3] M. Bazzoni, F. Terenziani, A. Secchi, G. Cera, I. Jabin, G. de Leener, AUTHOR INDEX M. Luhmer, A. Arduini, Chem. Eur. J. 2020, 26, 3022 – 3025. PO048 POSTERS / SMALL MOLECULES 201 ARTUR BRZEZICKI1,2, DIFFERENTIATION AND IDENTIFICATION OF ENANTIOMERS PIOTR GARBACZ1 BY NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY WITH 1University of Warsaw, Faculty of Chemistry, Laboratory of Nuclear SUPPORT OF QUANTUM MECHANICAL COMPUTATIONS Magnetic Resonance Spectroscopy, Differentiation and identification of the chiral molecules are crucial are chiral building blocks of solifenacin succinate, the API used to Warsaw, Poland 2ZF Polpharma SA, Starogard parts of the characterization of active pharmaceutical ingredients treat overactive bladder. W used two CDAs: Mosher’s acid (3) and Gdański, Poland (API). Therefore, the development of methods of the characteriza- 1-(9-anthryl)-2,2,2-trifluoroethanol (4). In the racemic mixtures of tion of optical isomers impacts the pharmaceutical industry, and 1, the shifts of 13C NMR peaks due to the presence of the CDAs were NMR spectroscopy is one of the tools used for this purpose. 0.05 ppm using ( R)-3 and 0.10 ppm after application of ( R)-4. For compound (3 R)-2, 1H NMR shifts induced by CDA were of the order Discrimination of chiral molecules by NMR is based on indirect for 0.06 ppm. methods since, in normal conditions, the NMR spectra of both enantiomers are indistinguishable. These methods utilize the for- Quantum chemical computations were performed in the Dalton mation of diastereomers whose spectra manifest chirality-induced computer program for the same pairs of compounds as measured shifts of NMR lines. For instance, one can use chemical solvating experimentally using the density functional theory. Then, obtained agents (CSAs), chemical derivatizing agents (CDAs), complexes in computations, nuclear magnetic shielding was converted to with metals, and chiral liquid crystals to differentiate enantiomers chemical shifts using data from Ref. [4]. Our final results show that [1−3]. The observed peak shifts may be attributed to the presence of the small amplitude of the induced chiral shifts makes it necessary a given enantiomer using empirical rules; however, a much more to model the dynamic interaction between studied compounds INTRODUCTION effective and general method would apply results of quantum me- and CDA very accurately since intermolecular interactions may af- SPONSORS chanical computations. fect the observed shifts in a degree comparable with that induced PROGRAM by CDA. We tested the latter approach using 1-phenyl-1,2,3,4-tetrahydroiso- PROGRAM — BY DAY quinoline (1) and 1-azabicyclo[2.2.2]octan-3-ol (2). The compounds TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] J. M. Seco, E. Quiñoá, R. Riguera. The Assignment of the Absolute Configuration by NMR Using Chiral Derivatizing Agents : A Practical Guide. Oxford University Press; 2015. POSTERS [2] T. J. Wenzel, Discrimination of Chiral Compounds Using NMR Spectroscopy. Wiley; 2007. [3] T. J. Wenzel, Strategies for using NMR spectroscopy to determine absolute configuration. ADVERTISERS Tetrahedron: Asymmetry. 2017; 28(10); 1212-1219. [4] W. Makulski, K. Jackowski, Journal of Magnetic Resonance 2020, 313, 106716. AUTHOR INDEX Acknowledgements: Calculations have been carried out using resources provided by Wroclaw Centre for Networking and Supercomputing (http://wcss.pl), grant No. 542. PO049 POSTERS / SMALL MOLECULES 202 LUCIA EMA SEKULA, RAPID CHARACTERIZATION OF IVA HABINOVEC, TETRAHYDROFURFURYL 3-MACROZONE DERIVATIVE IVANA MIKULANDRA, EMA HOŠNJAK, REACTION MIXTURE COMPONENTS USING LC-SPE/ PREDRAG NOVAK NMR APPROACH Department of Chemistry, Faculty of Science, Horvatovac 102a, Macrozones are novel bioactive conjugates of azithromycin and thiosemicarbazones that possess very good in vitro 10000 Zagreb, Croatia antibacterial activity against selected Gram-positive and Gram-negative bacteria [1]. Due to a global problem of growing bacterial resistance, discovery of new antibiotics is of an outmost importance and various studies are in progress to overcome the resistance mechanisms [2-3]. Efficient and rapid isolation, as well as structural characterization, of newly synthesized compounds are crucial steps prior to biological evaluation. Furthermore, impurity profiling is very important procedure during the drug development in pharmaceutical industry. Classical purification methods, such as preparative and semi-preparative liquid chromatography, can be time and solvent consuming. Using hyphenated systems enables shorter analysis time and reduces solvent consumption [4-5]. Hyphenated LC-SPE/NMR system enables efficient one step separation of reaction mixture components, extraction of each individual component on SPE-cartridge and structural characterization of isolated compounds by NMR spectroscopy. In this study we have successfully isolated and structurally characterized the newly synthesized tet- INTRODUCTION rahydrofurfuryl 3-macrozone derivative and main reaction mixture components using the LC-SPE/NMR methodol- SPONSORS ogy. Structures of the isolated compounds were readily elucidated on the basis of one- and two-dimensional NMR PROGRAM spectra and MS spectra. Some of the isolated components were diastereomers which were additionaly analysed by PROGRAM — BY DAY NOESY spectroscopy. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] I. Grgičević, I. Mikulandra, M. Bukvić, M. Banjanac, V. Radovanović, I. Habinovec, B. Bertoša, P. Novak, Int. J. Antimicrob. Agents 2020, 106147, INVITED AND PROMOTED LECTURES 5. [2] M. Bukvić Krajačić, M. Dumić, P. Novak, M. Cindrić, S. Koštrun, A. Fajdetić, S. Alihodžić, K. Brajša, N. Kujundžić, Bioorg. Med. Chem. Lett. 2011, 21, 853-856. POSTERS [3] B. Arsić, P. Novak, M.G. Rimoli, J. Barber, G. Kragol, F. Sodano, Macrolides: Properties, Synthesis and Applications, De Gruyter, Berlin, 2018. [4] I. Habinovec, ADVERTISERS T. Jednačak, P. Novak, ADMET & DMPK 2015, 3, 352-358. [5] M. Harča, I. Habinovec, E. Meštrović, I. Biljan, P. Novak, Croat. Chem. Acta 2016, 89, 543-547. AUTHOR INDEX Acknowledgements: This work was supported by the Croatian Science Foundation under the project IP-2018-01.8098, the Macrozones. PO050 POSTERS / SMALL MOLECULES 203 KRISTINA KRISTINAITYTE1, ILLUMINATED-NMR SPECTROSCOPY FOR THE MONITORING TOMASZ RATAJCZYK1, OF THE PHOTODIMERIZATION PROCESSES OF EXEMPLARY MARIUSZ PIETRZAK1, MAREK CZARNOTA2 ANTHRACENE DERIVATIVES 1Institute of Physical Chemistry, Polish Academy of Science, Warsaw, Poland It has been recently demonstrated that Illuminated Nuclear Magnet- faster compared to 9-bromoanthracene. However, the experimen- 2University of Warsaw, Faculty of Physics, ic Resonance (INMR) spectroscopy can be a very powerful tool for in- tal results showed that the reaction rate of the photodimerization Warsaw, Poland vestigation of photo-driven systems, as this method offers in-depth strongly depends on the experimental set-up. Therefore, a mixture insight into molecular structures, as well as different chemical proof anthracene and 9-bromoanthracene was also investigated. Thus, cesses at the molecular level [1]. Photochemical reactions of anthra- in the one sample with two compounds the photoreactions were cenes have long provoked interest due to their numerous applica- monitored under the same conditions. Estimated reaction rates tions. This class of aromatic organic compounds plays an extremely showed that the this time photodimerization of anthracene was important role as multi-faceted organic building blocks in nearly all about 2 times faster compared to 9-bromoanthracene. Moreover, aspects of chemical and industrial research [2], especially in the hi-in the system with two substrates, the formation of three possible tech and green energy sectors. Therefore, the goal of this work was to products was demonstrated. implement the INMR for the monitoring of their photodimerization. The presented approach helped us to evaluate the experimental Anthracene and its derivative 9-bromoanthracene were chosen as conditions and their influence on the photochemical reactions of the model compounds. Unless stated otherwise, NMR experiments anthracenes, and suggested that the relative reactivity obtained were performed at 9.4 T magnetic field after illuminating the sam-from the system with two substrates is more informative when the INTRODUCTION ples with a 365 nm wavelength LED. Dichloromethane was chosen comparison of several compounds is a goal. This will result in a bet- SPONSORS as a solvent. Different experimental conditions, such as the oxygen- ter understanding of the photochemical processes, which is essen- PROGRAM ation level, the method of sample preparation, the concentration, tial for the practical utilization of the photodimerization of acenes. PROGRAM — BY DAY as well as the volume of the illuminated sample were investigated, Moreover, the approach can be easily extended to different photoac- TOPICS and the effect of these conditions on photoreactivity was evaluated. tive reactions and processes. The obtained knowledge opens up an PRIZE LECTURES easy and reliable route for further development of the in-situ illumi- The analysis of reaction curves derived from 1H-NMR spectra re- PLENARY LECTURES nated-NMR monitoring system. vealed that the photodimerization of anthracene was about 10 times TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] A. Seegerer, Ph. Nitschke, R.M. Gschwind, Angew. Chem. Int. Ed. 2018, 57, 7493 –7497. AUTHOR INDEX [2] M. Yoshizawa, J.K. Klostermanb, Chem. Soc. Rev. 2014, 43, 1885-1898. PO051 POSTERS / SMALL MOLECULES 204 DAMJAN MAKUC1, NMR CHARACTERIZATION OF POLYSACCHARIDE FAZILET GÜRER2, DERIVATIVE SCAFFOLDS FOR REGENERATIVE TAMILSELVAN MOHAN3, 2, KARIN STANA KLEINSCHEK3, 4, MEDICINE JANEZ PLAVEC1,5,6 1 Nature itself can built multicomplex but still spatially resolved stable structures as shown on this image of bacterial National Institute of Chemistry, Slovenian NMR Centre, Ljubljana, Slovenia cell wall and knee cartilage. Idea of our colleagues from Maribor and Graz that design new biomaterials is to build 2University of Maribor, Faculty of Mechanical structures close to nature, using bioprinting and materials which are already present in living organism. Such bioEngineering, Laboratory for Characterisation materials can then be used for bone tissue regeneration. and Processing of Polymers, Maribor, Slovenia 3Graz University of Technology, Institute of In this study we characterized several polysaccharide-amino acid conjugates. Two different types of amino acid es-Chemistry and Technology of Biobased System (IBioSys), Graz, Austria ters: glycine- and tryptophan- were conjugated to the polysaccharide carboxymethylcellulose (CMC) in water using 4University of Maribor, Faculty of Electrical carbodiimide at ambient conditions. Engineering and Computer Science, Institute of Automation, Maribor, Slovenia Characterization of both CMC-amino acid conjugates is based on NMR spectra, which includes 1H, 13C-DEPT 135 5EN-FIST Centre of Excellence, Ljubljana, Slovenia as well as two-dimensional 1H-13C HSQC/HMBC correlation spectroscopy. Besides structure characterization there 6University of Ljubljana, Faculty of Chemistry were two questions that we tried to answer: can we provide solid and direct evidence for the successful conjugation and Chemical Technology, Ljubljana, Slovenia of the amino acid esters to the CMC backbone via an amide bond? Long-range proton-carbon correlation signals in the HMBC of the CMC-Gly-OMe between allowed unambiguous identification of the carboxyl ester group. Furthermore, the methylene group of the glycine moiety showed cor- INTRODUCTION relation signals to both carboxyl ester and carboxamide carbons. The second goal was to determine the degree of SPONSORS substitution, which means the amount of Trp-OMe that was found in Trp-OMe conjugates. The example here shows PROGRAM quantification of all functional groups, which were unambiguously assigned. Calculation confirms that the ratio PROGRAM — BY DAY between tryptophan and hyaluronic acid scaffold is approximately 1:3. TOPICS NMR characaterization of novel polysaccharide derivatives is an important step in further design and development PRIZE LECTURES of materials used for regenerative medicine. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] T. Mohan, A. Dobaj Stiglic, M. Beaumont, J. Konnerth, F. Gürer, D. Makuc, U. Maver, L. Gradišnik, J. Plavec, R. Kargl and K. Stana Kleinschek, AUTHOR INDEX ACS Appl. Bio Mater. 2020, 3 (2), 1197-1209. PO052 POSTERS / SMALL MOLECULES 205 WŁODZIMIERZ MAKULSKI, DETERMINATION OF SULPHUR-33 NUCLAR MAGNETIC MOMENT PIOTR GARBACZ FROM GAS-PHASE NMR STUDIES OF 3HE AND SF6 University of Warsaw, Faculty of Chemistry, Warsaw, Poland Gas-phase NMR spectroscopy can provide highly nucleus spin extrapolated shielding constant nuclear magnetic accurate and precise values of nuclear magnetic NMR frequency (ppm) moment dipole moments. Results of measurements con- (MHz) (nuclear magneton) ducted in condensed phases require corrections for nuclear magnetic shielding, bulk magnetic suscep- 33S 3/2 38.4197875(6) 392.6 (rel.) 0.6432555(10) tibility, and intermolecular interactions. The lat- 350.0 (non rel.) 0.6432281(10) ter factor may be omitted in the gas-phase studies since extrapolating the resonance frequency to the 3He 1/2 381.3575177(5) 59.96743(10) 2.127625308(10) zero-pressure limit removes the intermolecular ef- 19 fects [1]. F 1/2 471.0653234(5) 158.3 (rel.) 2.628321(13) We examined the density-dependences of 19F, 33S, 157.8 (non rel.) and 3He resonance frequencies in gaseous mixtures of helium and sulfur hexafluoride. The gaseous sam- At the constant external magnetic field B = 11.75 T, the measured resonance fre- ples contained small amounts of 3He in excess of SF 0 6 quencies are linearly dependent on SF densities. New experimental value μ(33S) cal- used as a buffer gas. As the reference nucleus, which 6 culated against that of 3He is +0.6430416(10) μ without shielding corrections and NMR parameters are known with high precision, N INTRODUCTION +0.6432555(10) μ with these corrections calculated in relativistic approach. This helium-3 was used in this study [2]. The total den- N SPONSORS more accurate result generally agrees with previously reported results [1]. Recalcu- sities of the samples were from 0.22 to 1.00 mol/L. PROGRAM lated from these data isotropic 19F shielding in SF molecule is σ (19F) = 406.2 ppm. The 19F, 33S, and 3He NMR resonance frequencies were 6 0 PROGRAM — BY DAY It is significantly higher than the absolute shielding constant obtained from the measured using the Varian INOVA 500 NMR spec- TOPICS relativistic theoretical approach. It means the necessity of new measurements 19F trometer at T = 300 K. They are collected in the table PRIZE LECTURES nuclear moment corrected by relativistic factors. below (for shielding constants, see ref. [3]). PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] A. Antušek, K. Jackowski, M. Jaszuński, W. Makulski, M. Wilczek, Chem.Phys.Lett. , 2005, 411, 111. [2] N.J. Stone, J. Phys. Chem. Ref. Data, 2015, 44, 031215. [3] K. Ruud, T. B. AUTHOR INDEX Demissie, M. Jaszuński, J. Chem. Phys. , 2014, 140, 194308. PO053 POSTERS / SMALL MOLECULES 206 BRUNO ALEKSANDER A SIMPLE FLOW CHART DIAGRAM BASED ON 1H–15N MARTEK, HMBC NMR SPECTROSCOPY FOR DISTINGUISHING MATEJA MIHELAČ, MARTIN GAZVODA, BETWEEN ISOMERIC AZAINDOLES IN NEW ILLEGAL JANEZ KOŠMRLJ DRUGS University of Ljubljana, Faculty of Chemistry and Chemical Recently, the development of new synthetic cannabinoid receptor agonists (SCARs), which are classified as psycho-Technology, Ljubljana, Slovenia active substances, has greatly progressed. However, their evolvement represents new challenges in terms of their detection and identification. Due to the lack of data, distinguishing between SCARs of isomeric nitrogen containing heterocyclic structure is difficult and represents one of the main obstacles [1]. The use of NMR spectroscopy, which is a corner-stone method for the identification and characterization of organic compounds, is of great help in solving this problem [2]. In particular, for the nitrogen containing compound, 1H– 15N HMBC spectroscopy plays an important role in structural identification [3]. Herein, a simple and rapid tool, based on 1H and 1H– 15N HMBC NMR spectroscopy for identification of different isomeric azaindole type of SCARs, is presented. The constructed flow chart diagram was also tested on seized sample, 5F-MDMB-P7AICA. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] B. A. Martek, M. Mihelač, M. Gazvoda, M. Virant, D. Urankar, M. Krivec, M. Gostič, B. Nemec, B. Koštrun, M. Janežič, S. Klemenc, J. Košmrlj, POSTERS Drug Test. Anal. 2019, 617. [2] Q. Liu, X. Zhu, Gen. Chem. 2015, 1, 26. [3] M. Köck, J. Junker, T. Lindel, Org. Lett. 1999, 1, 2041. ADVERTISERS Acknowledgements: This work was supported by grants from the Slovenian Research Agency (Research Core Funding Grant P1-0230 and projects J1-8147, AUTHOR INDEX J1-9166 and L1-9191) and by the National Forensic Laboratory, Ministry of the interior Police contracts C1714-17-460078 and C1714-19-460155. PO054 POSTERS / SMALL MOLECULES 207 MATEJA MIHELAČ, A RAPID DETERMINATION OF AZAINDOLE CORE IN BRUNO ALEKSANDER NEW ILLEGAL SUBSTANCES BY 1H–15N HMBC NMR MARTEK, MARTIN GAZVODA, CORRELATION SPECTROSCOPY JANEZ KOŠMRLJ One of the most important techniques for the analysis of the natural occurring and synthetic compounds is NMR University of Ljubljana, Faculty of Chemistry and Chemical Technology, (nuclear magnetic resonance) spectroscopy [1]. Various experiments using 2D NMR techniques such as COSY, HSQC Ljubljana, Slovenia and HMBC enable us to identify unknown complex compounds and determine their structure. 2D NMR techniques in addition to proton–carbon correlations, also allow us to determine the proton–nitrogen framework [2]. A rapid growth in the number of new synthetic cannabinoid receptor agonists (SCARs) renders this group of new psychoactive substances particularly demanding in terms of detection, identification, and responding. With no reference data available, differentiation and structural elucidation of constitutional isomers represents one of the major challenges. Since nitrogen is a common element in biological organic compounds, 1H–15N HMBC plays an important role in structural identification [3]. In determining the structure of nitrogen containing compounds, the use of 1H and 1H–15N HMBC NMR is inevitable, but sometimes interpretation of results can be time consuming. For this purpose, we constructed a simple flow chart diagram based on 1H and 1H–15N HMBC NMR spectroscopy for rapid and efficient determination of azaindole core. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Fukushi, E. Biosci., Biotechnol., Biochem. 2006, 70, 1803. [2] J. Furrer, Chem. Commun. 2010, 46, 3396. [3] B. A. Martek, M. Mihelač, M. Gazvoda, INVITED AND PROMOTED LECTURES M. Virant, D. Urankar, M. Krivec, M. Gostič, B. Nemec, B. Koštrun, M. Janežič, S. Klemenc, J. Košmrlj, Drug Test. Anal. 2019, 617. POSTERS Acknowledgements: This work was supported by EU ISF-Police programmes (R Slovenia MoI-GPD-NFL project IP.SO5.1.6-02), grants from the Slovenian ADVERTISERS Research Agency (Research Core Funding Grant P1-0230 and projects J1-8147, J1-9166 and L1-9191) and by the National Forensic Laboratory, Ministry of the AUTHOR INDEX interior Police contract C1714-17-460078 and C1714-19-460155. PO055 POSTERS / SMALL MOLECULES 208 MARKUS OBKIRCHER, ASSURE ACCURACY OF YOUR QUANTITATIVE NMR RESULTS ALEXANDER RUECK, BY qNMR STANDARDS, PERFORMANCE QUALIFICATION AND CHRISTINE HELLRIEGEL, ROMANA RIGGER PROFICIENCY TESTING Merck KGaA, Sigma-Aldrich, Buchs, Switzerland Since quantitative NMR (qNMR) spectroscopy is considered a rela- Second, it has to be demonstrated especially in regulated environ- tive primary method [1-3], the technology is becoming increasing- ments that the NMR instrument itself performs according to re- ly popular for the characterization and potency determination of quirements given by the authorities or guidelines. Through a col-organic compounds. In comparison to LC or GC applications the laboration between Bruker and Merck a two-component mixture method has a variety of advantages as no substance specific calibra-has been designed for its use as quantitative performance qualifi- tion is needed. However, in order to achieve accurate results three cation (qPQ) sample. The mixture consists of two CRM for qNMR in factors are of highest importance, which will be described in detail DMSO-d6 delivered in ampules and can be applied in combination in this presentation: with Bruker’s Assure software [8]. First, the method depends on the availability of suitable standards The last important factor is the proficiency of the personal operate.g. Certified Reference Material (CRM). CRM are characterized by ing NMR instruments and performing experiments. To assure high specific requirements such as traceability to the SI unit, measure-quality and accuracy of measurement results these individuals have ment uncertainty, homogeneity assessment, stability testing and a to demonstrate their expertise in regular intervals. For this purpose, hard expiry date. All these attributes are defined in ISO/IEC 17025 we developed a qNMR proficiency testing scheme, through which and ISO 17034 [4,5]. Through the last years, Merck has been working participants can compare their skills by reporting back their results INTRODUCTION on the development of neat CRM for use in 1H, 31P and 19F-qNMR as against a validated mean value. SPONSORS well as pre-dissolved ready-to-use standards [6,7]. The implementation of these three pillars – qNMR CRM standards, PROGRAM performance qualification and proficiency testing – assures preci- PROGRAM — BY DAY sion and accuracy of quantitative NMR measurements. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Malz F, Jancke H, Journal of Pharmaceutical and Biomedical Analysis 2005, 38(5), 813-823. [2] Saito T, Ihara T, Koike M, Kinugasa S, Fujimine Y, Nose K, Hirai T, Accreditation and Quality Assurance 2009, 14(2), 79-86. [3] De Bievre P, Dybkaer R, Fajgelj A, Hibbert BD, Pure and Applied Chemistry 2011, 83(10), 1873-1935. [4] ISO/IEC 17025:2005, “General requirements for POSTERS the competence of testing and calibration laboratories”. [5] ISO 17034:2016, “General requirements for the competence of reference material producers”. [6] Weber M, Hellriegel C, Rueck A, ADVERTISERS Wuethrich J, Jenks P, Obkircher M, Analytical and Bioanalytical Chemistry 2015, 407(11), 3115-3123. [7] Rigger R, Hellriegel C, Rueck A, Sauermoser R, Morf F, Breitruck K, Obkircher M, Journal AUTHOR INDEX of AOAC International 2017, 100(5), 1365-1375. [8] https://www.bruker.com/fileadmin/user_upload/8-PDF-Docs/MagneticResonance/NMR/brochures/nmr-pharma-qPQ. PO056 POSTERS / SMALL MOLECULES 209 ZUZANA OSIFOVÁ, HYDROGEN BONDING OF METHYLATED NUCLEOBASE ONDŘEJ SOCHA, ANALOGUES STUDIED BY NMR SPECTROSCOPY LUCIE MUŽÍKOVÁ-ČECHOVÁ, MICHAL ŠÁLA, Hydrogen bonding (H-bonding) is a type of weak noncovalent in- considered absent in human genome until 2018 [7]. Nowaday, this ZLATKO JANEBA, teraction with enormous importance for the life on Earth. They de- modification is related to development of obesity and Alzheimer’s MARTIN DRAČÍNSKÝ termine, for example, the structure and proper function of proteins disease. The methyl group in position 6 can adopt two orientations Institute of Organic Chemistry and and nucleic acids, however, they are an order of magnitude weaker because the bond between (methyl)amino group and purine ring is Biochemistry, Czech Academy of Sciences, than covalent bonds. According to Grabowski there are three types of an order higher than one, and the rotation around this bond is Prague, Czech Republic of hydrogen bonds – weak (bond energy between 0.5–4 kcal/mol), restricted. Each of the conformation isomers (rotamers) has a differ-moderate (4–15 kcal/mol) and strong (15–60 kcal/mol) [1]. ent hydrogen bonding pattern, and the steric hindrance of methyl group discriminates Watson-Crick or Hoogsteen base-pairing site. The intermolecular binding in nucleic acids (NAs) was described by Watson and Crick in 1953 [2] and further extended by Karst Hoog- In this work we present the NMR and DFT study of intermolecular steen few years later [3]. In Hoogsteen base pairing is purine base hydrogen bonds of methylated adenine analogues with their com-flipped by 180°, and the nitrogen atom in position 7 is involved in plemental partner, thymine. We studied the binding interactions of hydrogen bond. This alternative pairing was found e.g. in DNA tri-adenine derivatives by low-temperature NMR in different solvents. plexes [4] and quadruplexes [5]. We found out that the N-methylation stabilizes Hoogsteen base pairing. Hoogsteen base pairing is also preferred for those adenine The methylation of nucleobases is one of the most important epi- derivatives which are able to form both (Watson-Crick and Hoog- genetic mechanism associated with aging, genomic imprinting steen) complex types via two H-bonds. INTRODUCTION and tumorigenesis [6]. Adenine methylation in position 6 has been SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Grabowski, S. J., J. Phys. Org. Chem.. 2004, 17, 18-31. [2] Crick, F. H. C.; Watson, J. D., Proc R Soc Lon Ser-A. 1954, 223, 80. [3] Hoogsteen, K., Acta Crystallogr. 1959, 12, 822-823. [4] INVITED AND PROMOTED LECTURES Gilbert, D. E.; Feigon, J., Curr Opin Struc Biol. 1999, 9, 305-314. [5] Sen, D.; Gilbert, W., Nature. 1988, 334, 364-366. [6] Liang, D.; Wang, H.; Song, W.; Xiong, X.; Zhang, X. H.; Hu, Z. P.; Guo, H. H.; POSTERS Yang, Z. J.; Zhai, S. D.; Zhang, L. H.; Ye, M.; Du, Q., Biochem Bioph Res Co. 2016, 480, 120-125. [7] Xiao, C. L.; Zhu, S.; He, M. H.; Chen, D.; Zhang, Q.; Chen, Y.; Yu, G. L.; Liu, J. B.; Xie, S. Q.; Luo, F.; ADVERTISERS Liang, Z.; Wang, D. P.; Bo, X. C.; Gu, X. F.; Wang, K.; Yan, G. R., Mol Cell. 2018, 71, 306-318. AUTHOR INDEX Acknowledgements: This work was supported by Czech Science Foundation (grant No. 20-01472S). PO057 POSTERS / SMALL MOLECULES 210 LEO SVENNINGSSON TENSORVIEW FOR MATLAB Lund University, Lund, Sweden TensorView for Matlab[1] is a molecular visualization tool with a focus on NMR tensors, which is a transformative work from professor Leonard Mueller’s TensorView, which is made for Mathematica.[2] Good visualization tools are needed for researchers and within undergraduate education to mediate some of the more spatial concepts within the field of NMR. In particular for tensor rotations, the lack of conventions can cause great confusion for the uninitiated. For that reason, a toolbox is included with a few common rotation schemes used within the scientific community. The graphical user interface version of TensorView for Matlab can be run without a Matlab license using Matlab runtime, and is available for free through github at: https://github.com/LeoSvenningsson/TensorViewforMatlab. A script version of TensorView for Matlab with all source files are also available that can easily be glued together with other software. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] L. Svenningsson, A. Ludwig, L. Mueller 2021, POSTERS https://github.com/LeoSvenningsson/TensorViewforMatlab, mathworks.com/matlabcentral/fileexchange/55231-molecule3d. ADVERTISERS [2] L. Mueller, et al. Magn Reson Chem. 2019, 57, 211-223, AUTHOR INDEX https://sites.google.com/ucr.edu/Mueller/home/tensorview PO058 POSTERS / SMALL MOLECULES 211 MICHAEL J. TAYLOR1, INVESTIGATING THE ELUSIVE OXYGEN DONOR ANOKHI SHAH1,2, COPPER SITE WITHIN A PROTEIN TYPE SCAFFOLD SELLAMUTHU ANBU3, MARGAUX VERDU3, LUCY JENNINGS3, Figure: (a) Cartoon representation, rendered using ALLNOX, of IULIIA MIKULSKA4, models of the peptides studied in this work. The grey sphere is the SOFIA DIAZ-MORENO4, proposed metal binding site. The predicted distances between a HASSANE E. L. MKAMI1, metal in the engineered binding site and the nitroxide radical of GRAHAM M. SMITH1, the TOAC spin label, and the inter TOAC distances, are presented. JANET E. LOVETTA2, Four-pulse DEER experimental data for Gd(MB1-2 ) (blue) TOAC 3 ANNA F. A. PEACOCK3 and Cu(MB1-2 ) (orange) showing (b) the time domain data TOAC 3 after background correction and (c) the corresponding distance 1SUPA School of Physics and Astronomy, University of St Andrews, UK distributions. DEER experiments were performed at 10 K at Q-band 2BSRC, University of St Andrews, UK and 15 K at X-band. Data were processed using DEERAnalysis2019. 3School of Chemistry, University of Birmingham, Edgbaston, UK 4Diamond Light Source, Harwell Science and Innovation Campus, Didcot, UK Here we report on the preparation and characterisation of a highly elusive Cu(II) binding site bound exclusively to oxygen donor atoms within a miniature artificial protein scaffold, a parallel three-stranded coiled coil, through an oxygen-rich binding site. Though originally designed for binding of lanthanides1 (e.g. Gd(III)), the copper binding INTRODUCTION has been characterised by a range of spectroscopic techniques, and in particular EPR. EPR experiments with MB1-2 SPONSORS (Figure 1a) were able to unambiguously rule out nitrogen coordination as they found no evidence of either weakly PROGRAM coupled or directly bound nitrogen. The results were consistent with Cu(II) bound exclusively to oxygen donors, PROGRAM — BY DAY with evidence of directly bound water. EPR distance measurements using DEER and RIDME between bound Cu(II) TOPICS and nitroxide radical spin label MB1-2TOAC (Figure 1) further verified this assignment as comparison of the dis- PRIZE LECTURES tance distributions reveal that both Gd(III) and Cu(II) appear to be binding at the same site. Despite coppers preva- PLENARY LECTURES lence in biology and its affinity for oxygen donors, proteinaceous CuOx binding sites remain elusive. However, this TUTORIAL LECTURE technique demonstrates the power of a protein design approach to access chemistry not, to the best of our knowl- INVITED AND PROMOTED LECTURES edge, exploited by biology. POSTERS ADVERTISERS AUTHOR INDEX References: [1] Berwick, M. R, et al, J. Am. Chem. Soc. 2014, 136(4), 1166–1169. PO059 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 212 MAIDER BIZKARGUENAGA1, METABOLIC PROFILE OF PATIENTS RECOVERED FROM COVID-19 RUBEN GIL-REDONDO1, USING 1H-NMR METABOLOMICS CHIARA BRUZZONE1, TAMMO DIERCKS2, SARS-CoV-2 is a highly transmissible virus described for the first and the effect size and statistical difference of recovered vs. nega-ANA LAÍN1, time in Wuhan (China) in December 2019. COVID-19 dissease has an tives were summarized in forest plots, while data distribution and ITZIAR MARTÍN-RUÍZ3, incubation period from 1 to 14 days resulting in a plethora of symp- the probability density were represented by violin plots. JUAN ANGUITA3, toms and can cause even death in the most severe cases. On the NIEVES EMBADE1, Our preliminary results showed a decreased of ketone bodies aceto- metabolomic aspect of COVID-19, although there are several studies JOSÉ M. MATO1, acetic acid, 3-hydroxybutyric acid and acetone, which are produced which describe that this is a systemic infection whith a significant ÓSCAR MILLET1 predominantly in the liver from fatty acid oxidation-derived ace- impact on the metabolism [1], the question remains of to what ex- 1 tyl-CoA and are accumulated during acute phase of COVID-19 [1]. Precision Medicine and Metabolism Lab, tent the metabolism of patients recovered from this disease returns CIC bioGUNE, BRTA, Derio, Spain Also the synthesis of 2-hydroxybutyric acid, marker of oxidative 2 to normal values. NMR-based metabolomics, used in this study, is NMR Platform, CIC bioGUNE, BRTA, Derio, stress, was restored to normal values in recovered patients. More- Spain able to quantify and identify metabolites and lipoproteins at the 3 over, we observed a similar level of TG-VLDL and Apo-B/Apo-A1 Inflammation and Macrophage Plasticity same time [2]. The aim of this work was to determine whether there Lab, CIC bioGUNE, BRTA, Derio, Spain ratio in recovered patients vs. healthy donors that could be due to is a recovery of the metabolic and/or lipoprotein profile associated a reestablished hepatic capacity to oxidize acetyl-CoA in the mito-with patients diagnosed and recovered from COVID-19. chondria. However, the level of Glyc A, a maker related to a general We analyzed plasma samples from healthy donors (n=73) and pa- inflammation, is elevated as well as COVID-19 positive patients in tients that had been diagnosed with COVID-19 by a RT-PCR assay the acute phase of the disease [3]. and have recovered with at least 3 months of delay after the acute INTRODUCTION These results suggest that some of the changes that COVID-19 in- phase of the disease (n=70). The samples were measured in a 600 SPONSORS duces in the fatty acid metabolism of the SARS-CoV-2 positive pa- MHz IVDr spectrometer (Bruker Biospin, Germany), daily calibrated PROGRAM tients can be reversed to healthy values after their recovery. How- by following all the SOPs to ensure reproducibility and the highest PROGRAM — BY DAY ever, Glyc A inflammation marker is still increased after 3 months spectral quality. Three different 1H NMR experiments were recorded TOPICS of the SARS-COV-2 infection. Further studies are necessary using a per sample at 310 K: a (1D) 1H NOESY, a (1D) 1H CPMG and a (2D) JRe- PRIZE LECTURES larger cohort of recovered individuals from COVID-19. solved experiment. Individual metabolite or lipoprotein subclasses PLENARY LECTURES were quantified with Bruker’s B.I.Quant-PS and B.I.LISA methods TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] C. Bruzzone, et al., iScience 2020, 23, 101645. [2] N. Embade, et al., Sci Rep. 2019, 10;9(1):13067. [3] T. Kimhofer, et al., J. Proteome Res. 2020, 19, 4442−4454. PO060 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 213 THOMAS MAVROMOUSTAKOS1, ELENI CHONTZOPOULOU1, COMBINED NMR SPECTROSCOPY AND CHRISTINA PAPAEMMANOUIL2, MARIA V. COMPUTATIONAL STUDIES REVEAL NOVEL CHATZIATHANASIADOU2, DIMITRIOS KOLOKOURIS3, SOFIA KIRIAKIDI1, ATHINA KONSTANTINIDI3, IOANNA LOX INHIBITOR GEROGIANNI4/5, THEODORE TSELIOS4, IOANNIS K. KOSTAKIS3, EVANGELIA D. CHRYSINA5, DIMITRA HADJIPAVLOU-LITINA6, Lipoxygenase (LOX) is an enzyme playing a key-role in the inflammation pathway. LOX’s DEMETER TZELI1, ANDREAS G. TZAKOS2. metabolites are linked not only to inflammation but also to a variety of different diseases 1 such as asthma, atherosclerosis, rheumatoid arthritis, psoriasis, brain disorders and can- Department of Chemistry, National and Kapodistrian University of Athens, Panepistimioupolis Zografou, Greece cer [1]. Recently, the concept of “drug repurposing” has gained ground since it 2Department of Chemistry, Section of Organic Chemistry and Biochemistry, University of Ioannina, promotes the discovery of novel uses for approved drugs as a mean to deliver the Ioannina, Greece 3 quickest possible transition from bench to bedside [2] . Food supplements have been Department of Pharmacy, National and Kapodistrian University of Athens, Panepistimioupolis- Zografou, Greece also explored for their potent pharmaceutical properties [3]. The discovery of special 4Department of Chemistry, University of Patras, Rion, Greece properties to such compounds may accelerate their commercial circulation, since 5Institute of Biology, Medicinal Chemistry and Biotechnology, National Hellenic Research Foundation, Athens, Greece they have already been assessed for their safety and toxicity. In this investigation, NMR 6Department of Pharmaceutical Chemistry, School of Pharmacy, Faculty of Health Sciences, experiments in combination with in silico studies have been performed in order to explore Aristotle University of Thessaloniki, Thessaloniki, Greece the potency of aspartame to serve as an anti-inflammatory drug targeting LOX enzyme. Mo- lecular docking studies indicated that aspartame presents strong binding to LOX’s active site, while the stability of “LOX-aspartame” complex was further evaluated by molecular dynamics simulations. The molecular profile of aspartame inside the enzyme’s cavity was evaluated by Saturation Transfer Difference (STD) NMR experiments, where all the interac- INTRODUCTION tions that aspartame forms with the residues of LOX’s cavity have been discovered. These SPONSORS critical interactions unveiled by STD NMR have been confirmed by Quantum Mechanics PROGRAM calculations. Furthermore, in vitro biological assays indicated the compound’s inhibitory PROGRAM — BY DAY activity against LOX and pinpoint aspartame as a significant compound that could lead the TOPICS anti-LOX drug design process. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Mashima, R.; Okuyama, T. Redox Biol. 2015, 6, 297–310. [2] Oprea, T.I.; Bauman, J.E.; Bologa, C.G.; Buranda, POSTERS T.; Chigaev, A.; Edwards, B.S.; Jarvik, J.W.; Gresham, H.D.; Haynes, M.K.; Hjelle, B. et al. Drug Discov. Today Ther. Strateg. 2011, ADVERTISERS 8, 61–69. [3] US Food & Drug Administration Additional Information about High-Intensity Sweeteners Permitted for Use in AUTHOR INDEX Food in the United States. PO061 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 214 KATJA DOLENC1, INTERACTIONS OF THE SARS-CoV-2 Nsp3 SUD MAJA MARUŠIČ2, M DOMAIN WITH 3’-UTR G-QUADRUPLEXES OF HOST JANEZ PLAVEC1,2,3 1University of Ljubljana, Faculty mRNA of Chemistry and Chemical Technology, Ljubljana, Slovenia The present work focuses on the discovery of potential interactions of the non-structural protein 3 (Nsp3) of the 2Slovenian NMR Centre, National SARS-CoV-2 with four-stranded DNA structures – G-quadruplexes –found in human mRNA 3’-untranslated regions Institute of Chemistry, Ljubljana, Slovenia (3’-UTRs). Nsp3 consists of 1946 amino acid residues, which makes it the largest viral non-structural protein in 3EN-FIST Centre of Excellence, SARS-CoV-2. It is comprised of 8 domains, including three SARS unique domains (SUDs) [1,2]. The roles of several Ljubljana, Slovenia. domains, including SUD M, remain unknown, but there is evidence that Nsp3 associates with Nsp4 andNsp6 to form a molecular pore complex in double membrane vesicles that function as replication factories for the virus in the host cell [3]. It has also been shown that the absence of SUD M abolishes viral replication in vivo. It has been previously reported that SARS-CoV SUD M binds G-quadruplexes. Comparison of the genomic RNA sequences of SARS-CoV and SARS-CoV-2 show sequence homology in the putative binding region of SUD M [4,5]. Therefore, we hypothesize that the same domain inSARS-CoV-2 will also bind G-quadruplexes. Because the SARS-CoV-2 genomic RNA contains no classical G-quadruplex-forming sequences, we searched for interactions with host RNA instead, focusing on potential G-quadruplex-forming sequences found in the 3’-UTR mRNA transcripts of human genes coding for the mitogen MAPK 1, TAB 3 protein involved in NF-κB signalling, the pro-apoptotic protein Bbc3, and the RAS oncogene family RAB6B. Folding of the selected G-rich sequences was evaluated in the presence of potas- sium cations alone, and in the presence of a peptide originating from the SUD M domain of Nsp3. We showed that INTRODUCTION the peptide interacts with several of G-rich oligonucleotides and induces the formation of distinct G-quadruplex SPONSORS structures. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] V. B. O’Leary, O. J. Dolly, C. Höschl, , M. Černa, S. V. Ovsepian, Int. J. Mol. Sci. 2021, 22, 1–14. [2] P. V’kovski, A. Kratzel, S. Steiner, H. Stalder, V. ADVERTISERS Thiel, Nat. Rev. Microbiol. 2021, 19, 155–170. [3] M. Santerre, S. P. Arjona, C. N. Allen, N. Shcherbik, B. E.Sawaya, J. Neurol. 2020. [4] J. Tan et al. PLoS Pathog. 2009, AUTHOR INDEX 5, e1000428. [5] Y. Kusov, J. Tan, E .Alvarez, L. Enjuanes, R. Hilgenfeld, Virology 2015, 484, 313–322. PO062 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 215 MAJA KOLAR1,2, GUANINE RICH SEQUENCES IN GENOME OF MAJA MARUŠIČ2, SARS-CoV-2 JANEZ PLAVEC1,2,3 1University of Ljubljana, Faculty As the world has been shaken by the COVID-19 pandemic, the scientific world has seemingly never concentrated of Chemistry and Chemical on one single subject more than on the SARS-CoV-2 coronavirus. Regulatory regions of viral genomes often contain Technology, Ljubljana, Slovenia 2National Institute of Chemistry, guanine rich sequences that form structures called G-quadruplexes (G4s), with G4 ligands displaying antiviral prop-Slovenian NMR Centre, erties. G4s are four-stranded nucleic acid secondary structures that form through self-recognition of guanine bases. Ljubljana, Slovenia 3 G4s are involved in important cellular processes such as gene expression, telomere stabilization, transcription and EN-FIST Centre of Excellence, Ljubljana, Slovenia translation [1]. Guanine rich sequences of SARS-CoV-2 genome are therefore considered a promising therapeutic target [2]. The typical guanine rich sequences in the form of G N G N G N G , where N is any nucleotide, occur less 2–5 0–7 2–5 0–7 2–5 0–7 2–5 frequently in the SARS-Cov-2 genome compared to SARS-CoV [3]. This presents an opportunity to investigate atypical G-rich sequences with bulges and hairpin stem loops, which simultaneously represent additional sites for ligand binding. Moreover, hairpin stem loops were shown to stabilize the G4s and promote their folding [4,5]. Dual ligands that bind both duplex and G4 structures could also resolve common G4 ligands problems of non-selectivity and low specificity [6]. Many algorithms that find putative typical G4s exist, and our goal is to re-formulate chosen algorithms for atypical G4 detection to include both bulges and hairpin stem loops and characterize them in vitro with NMR spectroscopy and other biophysical techniques. INTRODUCTION We are re-formulating two chosen existing algorithms G4Hunter and QPARSE [7,8]. G4Hunter is a scoring algo- SPONSORS rithm that finds typical putative G4s and has an advantage of negatively scoring cytosine residues that would form PROGRAM duplexes with guanine residues, whereas QPARSE searches for degenerate potential G4s. It is a novel graph-based PROGRAM — BY DAY algorithm with a dynamic programming approach that includes search for hairpins and bulges, but only in the G4 TOPICS loop regions (N tracts), not in between G tracts. Reformulating these two algorithms with distinct advantages is thus PRIZE LECTURES our current approach on the way to find atypical G-quadruplexes in the genome of SARS-CoV-2. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] D. Varshney, J. Spiegel, K. Zyner, D. Tannahill, S. Balasubramanian, Nat. Rev. Mol. Cell Biol. 2020, 21, 459–474. [2] C. Zhao et al, Angew. Chem. POSTERS - Int. Ed. 2021, 60, 432–438. [3] H. Cui, L. Zhang, Front. Microbiol. 2020, 11, 567317. [4] T. Q. Ngoc Nguyen, K. W. Lim, A. T. Phan, Nucleic Acids Res. 2020, 48, ADVERTISERS 10567–10575. [5] T. Q. N. Nguyen, K. W. Lim, A. T. Phan, J. Phys. Chem. B. 2020, 124, 5122–5130. [6] T. Q. N. Nguyen, K. W. Lim, A. T. Phan, Sci. Rep. 2017, 7, 11969. AUTHOR INDEX [7] A. Bedrat, L. Lacroix, J. L. Mergny, Nucleic Acids Res. 2016, 44, 1746–1759. [8] M. Berselli, E. Lavezzo, S. Toppo, Bioinformatics, 2020, 36, 393–399. PO063 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 216 IVANA MIKULANDRA, INTERACTIONS OF AZITHROMYCIN AMINOPROPYL IVA KUŠEC, DERIVATIVES WITH E. COLI RIBOSOME STUDIED BY TOMISLAV JEDNAČAK, PREDRAG NOVAK NMR SPECTROSCOPY Department of Chemistry, Faculty of Science, Zagreb, Croatia Azithromycin is a semisynthetic macrolide antibiotic which possess excellent pharmacokinetic properties and demonstrates satisfactory biological effect. Macrolides are bacteriostatic antibiotics. They exert their biological activity by binding to the 50S subunit of the ribosome. Macrolides are effective against Gram-positive and some Gram-negative bacterial strains [1,2]. Due to the frequent and improper use of drugs bacterial resistance has emerged, so there is a need to discover more effective antibiotics. Azithromycin aminopropyl derivatives are precursors in synthesis of macrozones. Macrozones are novel bioactive macrolide derivatives and conjugates of azithromycin and thiosemicarbazones [3]. In order to fully understand the mechanism of macrozone bioactivity it is important to determine their bound conformation and interactions with biological targets such as ribosome [4]. In our study, we used NMR spectroscopy to analyse binding of azithromycin derivatives with ribosome isolated from Escherichia coli. Conformations in free and bound state were studied by NOESY (nuclear Overhauser effect spectroscopy) and trNOESY (transferred nuclear Overhauser effect spectroscopy) experiments. STD NMR (Saturation transfer difference Spectroscopy) was used to determine the binding epitopes of azithromycin derivatives. By these INTRODUCTION techniques relationship between structure and activity will be explored. SPONSORS These results should contribute to the discovery and design of new, more effective macrolide antibiotics. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] B. Arsić, P. Novak, J. Barber, M. G. Rimoli, G. Kragol, F. Sodano, Macrolides: properties, synthesis and applications, Walter de Gruyter, Berlin/ POSTERS Boston, 2018, 1–30. [2] A. Janas, P. Przybylski, Eur. J. Med. Chem. 2019, 182, 11166. [3] I. Grgičević, I. Mikulandra, M. Bukvić, M. Banjanac, V. Radovanović, I. ADVERTISERS Habinovec, B. Bertoša, P. Novak, Int. J. Antimicrob. Agents 2020, 5, 106147. [4] T. Jednačak, I. Mikulandra, P. Novak, Int. J. Mol. Sci. 2020, 21(20), 7799. AUTHOR INDEX Acknowledgements: This work was supported by the Croatian Science Foundation under the project IP-2018-01-8098, the Macrozones. PO064 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 217 YOLANDA PÉREZ1, SEMAPHORIN3A - GLYCOSAMINOGLYCANS INTERACTION ROMAN BONET2, AS TARGET FOR AXONAL REGENERATION MIRIAM CORREDOR2, CECILIA DOMINGO2, Semaphorin 3A (Sema3A) is a cell secreted protein that participates We present our recent work, in which the main puporse was to JORDI BUJONS2, in the axonal guidance pathways. In the adult central nervous sys- characterize the interaction of the whole Sema3A C-terminal poly- ÀNGEL MESSEGUER2 AND tem (CNS), Sema3A acts as a canonical repulsive axon guidance basic region (Sema3A 725-771) with GAGs and its inhibition. First, IGNACIO ALFONSO2 molecule, inhibiting CNS regenerative axonal growth and sprout- we produced, purified 15N,13C-labeled basic domain and performed 1NMR Facility and 2Department of ing. Therefore, interfering with Sema3A signaling is a therapeutic the backbone assignment by acquiring 3D 1H and 13C direct detected Biological Chemistry, Institute for target for achieving functional recovery after CNS injuries. Moreo- NMR experiments. The limited spectral dispersion, and the lack of Advanced Chemistry of Catalonia, Spanish National Research Council, ver, several studies have suggested a role for proteoglycans (PGs) defined secondary structure elements, predicted based on chemi-Barcelona, Spain in Sema3A and its Nrp1 receptor function. Some studies showed cal shifts, categorizes human Sema3A C-terminal polybasic region that Sema3A adheres to the PG component of the extracellular ma-as an intrinsically disordered region. Next, we used a combination trix (ECM) at perineuronal nets or cortical neurons, and selectively of biophysical techniques (NMR, SPR, Fluorescence and Heparin Af-binds to heparin and chondroitin sulfate-E (CS-E) glycosaminogly- finity Chromatography) to gain insight into the interaction of the can (GAG) [1]. In our previous work, we identified a peptoid, termed Sema3A C-terminal domain with GAGs. These analyses confirmed SICHI (semaphorin-induced chemorepulsion inhibitor), that blocks that Sema3A C-terminal polybasic region binds to GAGs, preferably Sema3A chemorepulsion and growth-cone collapse in axons at the to heparin, and allowed us to identify the specific residues involved extracellular level [2]. Later, we proposed that SICHI blocks the bioin the interaction. Last, we studied the effect of a new peptoid mol- logically relevant interaction between Sema3A and GAGs compet- ecule (CSIC002) in the ineraction between Sema3A and heparin INTRODUCTION ing with Sema3A C-terminal polybasic region for binding to GAGs (dp14 oligosaccharide). We observed a displacement of Sema3A ba- SPONSORS [3]. For these earlier studies, we used basic peptides derived for sic tail from heparin by CSIC002 using 2D NMR 1H,15N-HSQC spec- PROGRAM Sema3A basic tail. tra chemical shift pertubation. Our structural study paves the way PROGRAM — BY DAY toward the design of new molecules targeting these protein-GAG TOPICS interactions with potential therapeutic applications. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Dick, G., Tan, C. L., Alves, J. N., Ehlert, E. M., Miller, G. M., Hsieh-Wilson, L. C., Sugahara, K., Oosterhof, A., van Kuppevelt, T. H., Verhaagen, J., Fawcett, J. W., & Kwok, J. C. J. Biol. INVITED AND PROMOTED LECTURES Chem. 2013 288(38), 27384–27395 [2] Montolio, M., Messeguer, J., Masip, I., Guijarro, P., Gavin, R., Antonio Del Rio, J., Messeguer, A., & Soriano, E. Chem. Biol. 2009, 16(7), 691–701 [3] Corredor, POSTERS M., Bonet, R., Moure, A., Domingo, C., Bujons, J., Alfonso, I., Perez, Y., & Messeguer, A. Biophys. J. 2016, 110(6), 1291–1303. ADVERTISERS Acknowledgements: We acknowledge the financial support from the European Union Seventh Framework Programme (FP7/2007-2013) under the Project VISION, Grant Agreement n° AUTHOR INDEX 304884. PO065 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 218 YULIA PUSTOVALOVA, BACKBONE ASSIGNMENT OF Cov-Y DOMAIN FROM OKSANA GORBATYUK, NONSTRUCTURAL PROTEIN 3 OF SARS-Cov-2 BING HAO, YUNFENG LI, Like other (+) RNA viruses severe acute respiratory syndrome coro- sequence contains the most C-terminal 286 residues of nsp3 (1660- JEFFREY C. HOCH navirus 2 (SARS-Cov-2 or covid-19) forms specialized membra- 1945). It was expressed and purified as described [1] at a yield of University of Connecticut Health, nous organelles in infected cells. These double membrane vesicles 20mg of the protein from 1L deuterated M9 media with 15N-labeled Department of Molecular Biology (DMVs) made from ER membrane serve the purpose of creating fa- ammonium chloride and 13C-labeled glucose. All NMR experiments and Biophysics, Farmington CT, USA vorable microenvironment for viral replication machinery by con- were done with a sample of 0.35 mM 2H,13C,15N-labeled nsp3 Cov-Y in centrating viral components and at the same time shielding them 25 mM MOPS buffer pH 6.4 with 100 mM LiBr and 2 mM DTT. from the host immune defenses. Nonstructural protein 3 (nsp3) of Backbone assignment of nsp3 Cov-Y from SARS-Cov-2 were per-SARS-Cov-2 together with nsp4 and nsp6 are involved in DMV for- formed with TROSY-based triple resonance experiments: HNCO, mation. Nsp3 has 1945 residues that comprise more than a dozen HNCA, HNCACB, HNCACO and 15N-NOESY. All NMR experiments domains. The C-terminal part of SARS-Cov-1 nsp3, including two were acquired at 25°C using an Agilent 800MHz spectrometer transmembrane and three extramembrane domains, is necessary equipped with a cold probe. for DMV formation [1]. This region is highly conserved among all coronaviruses but there has been no structural information on any Nsp3 Cov-Y is a fully folded protein domain with small flexible re-C-terminal domains previously reported. Here we present an NMR gions at the termini. Nearly full 1H, 13C and 15N backbone assignment study of the most C-terminal domain of nsp3, Cov-Y. was obtained (96% HN, 94% N, 96% C, 96% Ca, 90% Cβ). Secondary structure elements were predicted by Talos+ based on measured We reported protein expression and purification protocol for nsp3 INTRODUCTION backbone chemical shifts. Nsp3 Cov-Y consists of 9 long (more than Cov-Y of SARS-Cov-2 in a collaborative article with the internation- SPONSORS 5 residues) a-helixes and 11 β-strands. The excellent NMR data qual- al covid-19 research consortium [2]. Here we present an optimized PROGRAM ity provides a firm basis for structure determination. construct with the first 22 flexible residues deleted. The final protein PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] M. M. Angelini et al., mBio. 2013, 4, e00524-13. [2] N. Altincekic et al., Front. Mol. Biosci. 2021, in press. ADVERTISERS Acknowledgements: This work is a contribution for the intervention Covid-19 NMR Consortium (https://covid19-nmr.de), and was supported by the US National Science Foundation via grant AUTHOR INDEX RAPID 2030601. PO066 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 219 MARCO SCHIAVINA1, THE DISORDERED REGIONS OF THE SARS-CoV2 LETIZIA PONTORIERO1, NUCLEOCAPSID PROTEIN STUDIED BY CARBON-13 VLADIMIR N. UVERSKY2, ISABELLA CATERINA FELLI1, NMR EXPERIMENTS WITHIN THE 1-248 CONSTRUCT ROBERTA PIERATTELLI1 1 The SARS-CoV2 nucleocapsid protein (N-protein) is a multifunctional and multidomain protein characterized by CERM and Department of Chemistry “Ugo Schiff”, University of Florence, Sesto three intrinsically disordered regions (IDRs), which cover about 35% of the primary sequence, separated by two Fiorentino, Florence, Italy. globular domains (NTD and CTD). Previous studies performed on the homologous protein from SARS-CoV demon- 2 Department of Molecular Medicine and strated that the three IDRs are involved in different processes, ranging from binding with RNA to interaction with USF Health Byrd Alzheimer's Research Institute, Morsani College of Medicine, other viral proteins [1]. High-resolution data that could describe the role of these IDRs are fundamental to access University of South Florida, Tampa, USA atomic information about the whole N-protein both in its native conditions and upon interactions with partners and targets. We started focusing on the NMR characterization of the first two IDRs of N-protein, the N-terminal linker (IDR1, 1-44) and the central serine-arginine rich region (IDR2, 181-248) in the 1-248 construct (IDR1-NTD-IDR2). A set of 13C detected experiments were acquired to assign the resonances of the disordered signals and to access atomic resolution for these two IDRs thanks to the high resolution that could be achieved with the carbon detection approach [2-3]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Chang, C. K.; et. al. Antiviral. Res. 2014, 103 (1), 39–50. [2] Felli, I. C.; Pierattelli, R., (Eds), 2015 “Intrinsically Disordered Proteins Studied by NMR INVITED AND PROMOTED LECTURES Spectroscopy”; Springer, Switzerland. [3] Schiavina, M.; et al.” Biomolecular NMR assignments 2021, l. 15,1 219-227. POSTERS Acknowledgements: The support and the use of resources of the CERM/CIRMMP centre of Instruct-ERIC is gratefully acknowledged. This work was supported ADVERTISERS in part by the Fondazione CR Firenze and by the Italian Ministry for University and Research. Fabio Almeida, Andreas Schlundt and Leonardo Gonnelli are AUTHOR INDEX acknowledged for the stimulating discussions. PO067 POSTERS / DRUG DESIGN AND COMBAT AGAINST COVID-19 220 T.MAVROMOUSTAKOS1, NUCLEAR MAGNETIC RESONANCE AND AB INITIO NIKOLETTA ZOUPANOU1, STUDIES OF ORGANOTIN METALLOTHERAPEUTICS S. CHADJIKAKOU2, C. BANTI2, This study involves the structure elucidation and conformational analysis of three metallotherapeutics organotin M.E STATHOPOULOU2, (IV) derivatives of cholic acid (CAH) with the formulae R Sn(CA) (R= Ph- (1), n-Bu- (2)) and R Sn(CA) (R= Me- (3). 3 2 2 S.KYRIAKIDI1, The structures of compounds (1-3) were determined using a combination of homonuclear and heteronuclear 2D CARLOS SILVA LOPEZ3 NMR spectroscopy. Subsequently, 119Sn NMR experiments and some semi-empirical quantum mechanics compu- 1National and Kapodistrian University of tations (method PM6) were performed, in order obtain the conformational properties and the possible geometry Greece, Department of Chemistry, Greece 2 of complexes under study. Using 119Sn chemical shifts of compounds (1-3) and comparing them to the reference University of Ioannina of Greece, Department of Chemistry, Greece compound SnCl , it was found that the geometry is triangular dipyramid for (1) and (2), and octahedral geometry 4 3University of Vigo of Spain, Department of for (3). Semi-empirical quantum mechanics computations PM6 [1], based on the Neglect of Diatomic Differential Organic Chemistry, Spain Overlap (NDDO), contributed to the estimate of an optimized conformation and the geometry of compounds too. According to the obtained results, their optimized geometry is distorted tetrahedral. Biological activity of these compounds is related with the conformation of atoms which surround the metal of Sn. Having received all in vitro results and obtaining the necessary information about the structure and the conformation of molecules, it was deemed necessary to perform in silico studies. This study was about the binding of compounds to the receptor 1A52 of ER-a (Estrogen receptor alpha ligand) hormone, to provide a plausible explanation and proceed to a rational design of new molecules. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] Stewart, J. J. P. J. Mol. Model. 2007, 13 (12), 1173– 1213. PO068 POSTERS / METABOLOMICS 221 RITA ARAÚJO1, TUMOR LIPID SIGNATURES DESCRIPTIVE OF ACQUISITION CAROLINE LAMB2, OF RESISTANCE TO ENDOCRINE THERAPY VICTORIA FABRIS2, CLAUDIA LANARI2, IN AN ENDOCRINE-RELATED BREAST CANCER MOUSE MODEL LUISA HELGUERO3, ANA M. GIL1 Breast cancer (BC) is the most common type of cancer in women to the acquisiton of hormonal independence and of ET-resistance, 1 and, in most cases, it is hormone-dependent (HD), thus relying on using NMR metabolomics. Tumor tissue and non-compromissed Department of Chemistry and CICECO - Aveiro Institute of Materials, steroid hormones (estrogens/progestins) to activate intracellular mammary gland obtained from mice implanted subcutaneously University of Aveiro, Portugal;2Instituto receptors and stimulate tumor growth [1]. Endocrine therapy (ET) with HD, HI and HIR tumors from the medroxyprogesterone ace-de Biología y Medicina Experimental aiming at hormone receptors is the primary treatment strategy, tate (MPA)-induced BC mouse model [4] were analyzed. Compared (IBYME), CONICET, Buenos Aires, Argentina;3iBIMED—Institute for however, about half of the subjects, develop resistance in time [2]. to healthy tissue, the tumors exhibited global increases in total/free Biomedicine, Department of Medical This involves the development of hormone independent (HI) tum- cholesterol, phosphatitylcholine (PtC), phosphatidylethanolamine Sciences, Universidade de Aveiro, Portugal ors, initially therapy-responsive and which subsequently become (PtE), sphingomyelin (SM), arachidonic/hexadecanoic acids (ARA/ resistant (HIR) to ET. The mechanisms that promote HI to HIR con- DHA), and decreases in monounsaturated and polyunsaturated fat- version are varied, and not completely understood. Lipid metabo- ty acids (MUFAs/PUFAs) and triacylglycerides (TAG). The HD to HI lism is affected by tumorigenesis and changes in lipid metabolism transition was characterised by increases in ARA/DHA and PtE and are associated with anti-cancer drug resistance [3]. Lipids can func-decreases in MUFAs/PUFAs and TAG. HI transition to HIR involved tion as lipid signalling molecules, influence the metastatic potential increases in ARA/DHA, free/total cholesterol, PtC, PtE and SM. Pu- of cancer cells, protect or enhance oxidative stress and are import- tative interpretation of these results is discussed and possible mark- ant mediators of the immune response [3]. Therefore, the aim of ers of resistance are suggested. INTRODUCTION this work was to characterize the specific lipid signatures associated SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] C. H. Yip, A and Rhodes, Future Oncol. 2014, 10, 2293-2301.[2] P. Augereau, A. Patsouris, E. Bourbouloux, C. Gourmelon, S. A. Lacourtoisie, et al., Ther. Adv. Med. Oncol. 2017, 9, 335-346. [3] N. Germain, M. Dhayer, M. Boileau, Q. Fovez, J. Kluza, Biology, 2020, 9, 1-21.[4] C. Lanari, C. A. Lamb, V. Fabris, L. Helguero, R. Soldati, et al ., Endocr. Relat. Cancer. 2009,16, 333-350. TUTORIAL LECTURE Acknowledgements: This work was developed within the scope of project CICECO-Aveiro Institute of Materials, FCT Ref. UID/CTM/50011/2019 and POCI-01-0145-FEDER-007679, financed INVITED AND PROMOTED LECTURES by national funds through the FCT/MCTES from the Foundation for Science and Technology (FCT). POSTERS We also acknowledge funds from FCT, within UID/BIM/04501/2019, POCI-01-0145-FEDER-007628 and pAGE (CENTRO-01-0145-FEDER-000003) projects, granted to the Institute ADVERTISERS of Biomedicine of Aveiro (iBiMED). Funding is also acknowledged from the Portuguese National NMR Network (RNRMN) supported by FCT and from through grant PD/BD/142850/2018) AUTHOR INDEX –PTNMR PhD programme. PO069 POSTERS / METABOLOMICS 222 DANIELA S. C. BISPO1, METABOLIC MARKERS OF OSTEOGENIC CATARINA S. H. JESUS1, DIFFERENTIATION OF HUMAN ADIPOSE-DERIVED LENKA MICHÁLKOVÁ2,3, MARIANA B. OLIVEIRA1, STEM CELLS THROUGH NMR METABOLOMICS JOÃO F. MANO1, ANA M. GIL1 Mesenchymal stem cells (MSCs - multipotent cells) can differentiate into a variety of cell lineages, namely osteo-1 genic, thus having great potential in bone regenerative medicine. Metabolomics offers exquisite insight into the Department of Chemistry, CICECO – Aveiro Institute of Materials, University metabolism of living organisms. However, only a few reports have monitored osteogenesis, with mass spectrom- of Aveiro, Campus Universitário de etry approaches predominating [1-3] compared to nuclear magnetic resonance (NMR) spectroscopy [4]. NMR fin- Santiago, Aveiro, Portugal 2 gerprinting and preliminary results on footprinting of human adipose tissue MSCs (hAMSCs) during osteogenesis Department of Analytical Chemistry, Institute of Chemical Process are presented, throughout the 21 days of osteogenesis in 2D cultures. Endometabolome results showed significant Fundamentals of the CAS, Prague, Czech dynamic differences in several amino acids, creatine/phosphocreatine, choline compounds, glycerolipids, phos- Republic 3Department of Analytical Chemistry, pholipids, among others. Moreover, preliminary results indicate alanine, glycerol and citrate as important secre-University of Chemistry and Technology, tome components. A clear metabolic separation was observed before and after 7 days of culture, although metabolic Prague, Czech Republic changes are observed from early on until day 21. This work paves the way to characterize the dynamic metabolism of stem cells during osteogenesis, ultimately enabling its monitoring through metabolic biomarkers, eventually translatable to in vivo clinical tissue regeneration strategies. INTRODUCTION SPONSORS References: [1] E. Alakpa, V. Jayawarna, A. Lampel, K. Burgess, C. West, S. Bakker, S. Roy, N. Javid, S. Fleming, S. Lamprou, J. Yang, A. Miller, A. Urquhart, P. PROGRAM Frederix, N. Hunt, B. Péault, R. Ulijn, M. Dalby, Chem. 2016, 1, 298. [2] M.Klontzas, S.Vernardis, M.Heliotis, E.Tsiridis, A.Mantalaris, Stem Cells Dev. 2017, 26, PROGRAM — BY DAY 723. [3] M.H.Amer, M.Alvarez-Paino, J.McLaren, F.Pappalardo, S.Trujillo, J.Q.Wong, S.Shrestha, S.Abdelrazig, L.A.Stevens, J.B.Lee, D.H.Kim, C.González-García, TOPICS D.Needham, M.Salmerón-Sánchez, K.M. Shakesheff, M.R. Alexander, C .Alexander, F.R. Rose, Biomaterials 2021, 266, 120450. [4] D. Gaur, Y.Yogalakshmi, S.Kulanthaivel, T.Agarwal, D.Mukherjee, A.Prince, A.Tiwari, T.Maiti, K.Pal, S.Giri, M.Saleem, I. Banerjee, Adv. Biosys. 2018, 2, 1800093. PRIZE LECTURES Acknowledgements: We acknowledge the Portuguese Foundation for Science and Technology (FCT) for co-funding the BIOIMPLANT project (PTDC/BTM- PLENARY LECTURES ORG/28835/2017) through the COMPETE2020 program and European Union fund FEDER (POCI-01-0145-FEDER-028835). CSHJ is grateful to the same TUTORIAL LECTURE project for funding their contracts with the University of Aveiro. DSCB acknowledges the Sociedade Portuguesa de Química and FCT for her PhD grant INVITED AND PROMOTED LECTURES SFRH/BD/150655/2020. LM would like to thank to the Operational Programme Research, Development and Education (EF18_053/0016920) provided by Ministry of Education, Youth and Sports of the Czech Republic. AMG acknowledges the CICECO-Aveiro Institute of Materials project, (UIDB/50011/2020 POSTERS & UIDP/50011/2020), financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership ADVERTISERS Agreement. The NMR spectrometer used in this work is part of the National NMR Network (PTNMR) and, partially supported by Infrastructure Project Nº AUTHOR INDEX 022161 (co-financed by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). PO070 POSTERS / METABOLOMICS 223 CHIARA BRUZZONE1, RUBÉN GIL-REDONDO1, A METABOLIC SYNDROME DEFINITION BY MARISA SECO2, ROCÍO BARRAGÁN3,4, URINE NMR-METABOLOMICS LAURA DE LA CRUZ1, TAMMO DIERCKS1, MAIDER BIZKARGUENAGA1, ANA LAÍN1, Metabolic syndrome (MetS) is a complex health condition that arise as a result of the pres- OSCAR COLTELL4,5, ELISABETTA BUGUIANESI6, ence of various risk factors: problems with the metabolism of the glucose, obesity, elevated NIEVES EMBADE1, QUENTIN M. ANSTEE7, levels of triglycerides, low HDL cholesterol and hypertension. Due to the increasing number DOLORES CORELLA3,4, JOSÉ M. MATO1, OSCAR MILLET1 of people affected by this desorder, MetS constitutes a huge worlwide problem [1] but un- 1CIC bioGUNE, BRTA, CIBERehd, Derio, Bizkaia, Spain fortunately, the molecular basis of MetS are not yet well understood. Several associations 2OSARTEN Kooperativa Elkartea, Arrasate-Mondragón, Spain 3 have tried to determine a medical definition for this syndrome but these diagnostic crite- Department of Preventive Medicine and Public Health, School of Medicine, University of Valencia, Valencia, Spain ria are only based on the compatible symptomatology. In this study we have analised by 4CIBER Fisiopatología de la Obesidad y Nutrición, Madrid, Spain NMR-metabolomics a large cohort of urine samples (from 11,754 individuals, 18-75 years 5Department of Computer Languages and Systems, Universitat Jaume I, Castellón, old) to examine the molecular signature of MetS. Volounteers were classified into 16 differ- Spain 6Gastroenterology Department, University of Turin, Turin, Italy ent profiles to populate all possible intermediate conditions from the total absence of any 7Translational & Clinical Research Institute, Faculty of Medical Sciences, Newcastle risk factor up to the presence of individuals with MetS (4-5%, depending on the definition). University, Newcastle-upon-Tyne, UK NMR metabolomics has shown to be sensitive to MetS, with each one of the contributing risk factors represented by at least one of the identified metabolite that have shown to be up- or down-regulated in this disease. This led to the design of a metabolic model of the syndrome that can discriminate between individuals with and without MetS with statistical significance. Aging and non-alcoholic fatty liver disease (NAFLD) are also risk factors that INTRODUCTION have been previosly associated with MetS [2,3], but our results evidenced that they do not SPONSORS directly interfere with the metabolic discrimination of the syndrome. In conclusion, thanks PROGRAM to the obtained results our model add an unprecedented diagnostic molecular dimension PROGRAM — BY DAY and may potentially improve clinical decision guiding early interventions in this syndrome. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] P.M. Nilsson, J. Tuomilehto, L. Rydén, Eur J Prev Cardiol. 2019, 26, 33–46. [2] H. Yki-Järvinen, Lancet Diabetes AUTHOR INDEX Endocrinol. 2014, 2, 901–10. [3] F. Bonomini, L.F. Rodella, R. Rezzani, Aging Dis. 2015, 6, 109–20. PO071 POSTERS / METABOLOMICS 224 TATIANA J. CARNEIRO1, NMR METABOLIC PROFILE OF THE IN VIVO IMPACT OF A RITA ARAÚJO1; POTENTIAL ANTICANCER DRUG - PD (SPERMINE) MARTIN VOJTEK2; 2 SALOMÉ GONÇALVES- Platinum (Pt(II))-containing drugs are currently some of the most The magnitude and dynamics of metabolome response to both MONTEIRO2; used antineoplastic agents in the treatment of several types of solid drugs are tissue-dependent, as expected, with kidney as the most CARMEN DINIZ2; tumors [1]. In particular, cisplatin (cDDP) is used in lung, testicular, affected organ, followed by liver and breast tissue. Polar metabolites ANA L. M. BASTISTA DE breast, and ovarian cancer therapies, although tumor acquired-re- tend to respond more strongly and rapidly (1 h) to Pd Spm than to 2 CARVALHO3; sistance and high toxicity are limiting factors for its clinical use [1]. cDDP in all tissues, with main variations in levels of amino acids, MARIA PAULA M. MARQUES3,4; Therefore, palladium (Pd(II))-complexes are increasingly emerging nucleotides, Krebs cycle intermediates ( e.g. succinate and fuma- ANA M. GIL1 as alternatives to Pt(II)-based drugs, mostly due to chemical sim- rate), and other metabolites such as dimethylamine and dimethyl 1Department of Chemistry and CICECO, ilarities to Pt(II) and initial promising anti-tumor properties [2]. sulfone. On the other hand, lipid metabolism exhibits a delayed (48 University of Aveiro, Aveiro, Portugal 2 In particular, Pd(II)-complexes with biogenic polyamines, such as h) or weak response in kidney and liver/ breast tissue, respectively. LAQV/REQUIMTE, Department of Drug Sciences, University of Porto, Porto, Portugal spermine (Pd Spm), have exhibited beneficial cytotoxic properties In brief, Pd Spm triggers faster responses and recovery to control 2 2 3R&D Unit “Molecular-Physical Chemistry”, [3,4]. Metabolomics can provide a holistic view of the metabolic levels for all organs tested, except for kidney lipophilic metabolism. University of Coimbra, Coimbra, Portugal 4 profile of drug-exposed tissues, giving useful insights into drug These results are suggestive of a potential lower adverse effects/ Department of Life Sciences, University of Coimbra, Coimbra, Portugal mechanisms and potentially revealing new markers of drug effica- toxicity of Pd Spm exposure, encouraging its potential use as an an- 2 cy and toxicity [5]. ticancer drug for clinical trials. This work reports a 1H NMR metabolomics characterization of the Pd Spm impact, compared to cDDP, on the metabolism of healthy 2 INTRODUCTION mice. Polar and lipophilic extracts were obtained from kidney, liver, SPONSORS and breast tissue of female BALB/c mice, excised at 1, 12 and 48 h PROGRAM after a single-dose injection of 2.7 mg/kg Pd Spm, 3.5 mg/kg cDDP, 2 PROGRAM — BY DAY and 200 μL of phosphate-buffered saline for the controls. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] T. Makovec, Radiol. Oncol. 2019, 53, 148-158. [2] M. P. M. Marques, Hindawi 2013, 2013, 287353. [3] T. J. Carneiro, A. S. Martins, M. P. M. Marques, A. M. Gil, Front. Oncol. 2020, 10, 590970. [4] T. J. Carneiro, R. Araújo, M. Vojtek, S. Gonçalves-Monteiro, C. Diniz, et al., Metabolites 2021, 17, 114. [5] T. J. Carneiro, R. Araújo, M. Vojtek, S. Gonçalves-Monteiro, C. Diniz, et INVITED AND PROMOTED LECTURES al., Metabolites 2019, 9, 279. POSTERS Acknowledgements: This work was developed within the scope of the project CICECO-Aveiro Institute of Materials, FCT Ref. UIDB/50011/2020 and UIDP/50011/2020, financed by national ADVERTISERS funds through the FCT/MCTES from the Foundation for Science and Technology (FCT) and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. We also AUTHOR INDEX acknowledge the Portuguese National NMR Network (RNRMN), supported by FCT funds. T.J.C. thanks FCT for her PhD grant SFRH/BD/145920/2019. PO072 POSTERS / METABOLOMICS 225 1 FRANCESCA DI CESARE 1, H NMR-BASED METABOLOMICS TO INVESTIGATE THE GHINI V.1, TENORI L. 3, EFFECT OF PROBIOTICS ON HUMAN PHENOTYPE PANE M. 4, AMORUSO A.4, SQUARZANTI D. 4, The human gut hosts around one-thousand of different species of commensal microorganisms that play a crucial AZZIMONTI B. 5,6, role in health promotion, implementing host’s physiology and metabolism [1]. In this perspective, probiotics are LUCHINAT C. 1,2,3 increasingly used, with the final aim of manipulating the composition of the gut microbiota, and with the aim of 1Magnetic Resonance Center (CERM), improving balanced microbial communities [2,3]. In this context, the challenge is to characterize and to under- University of Florence, Sesto Fiorentino, Italy 2 stand potential metabolic changes that could determine and affet the dynamic relationship between host and Consorzio Interuniversitario Risonanze Magnetiche di Metallo Proteine (CIRMMP), microbiome. Sesto Fiorentino, Italy 3Department of Chemistry, University of Nuclear Magnetic Resonance (NMR)-based metabolomics offers the possibility to investigate hundreds of various Florence, Sesto Fiorentino, Italy metabolites and lipids detectable in biological fluids ( i.e. serum, urine, etc.), providing a global representation of the 4Probiotical S.p.A., Novara, Italy 5Department of Health Sciences (DiSS), molecular mechanisms and potential effects of the dynamic and evolving interactions between the microflora and University of Piemonte Orientale (UPO), host, and of the response to probiotic assumption [4,5]. Novara, Italy 6Center for Translational Research on In this study, using a NMR-based metabolomic approach, we highlighted the molecular effects obtained by micro- Autoimmune and Allergic Diseases (CAAD), organisms modulation through probiotic treatment, on human urine and serum metabolome. Twenty-one healthy DiSS, UPO, Novara, Italy volunteers were enrolled in the study and administered with two different dosage of probiotic (high and low) for a total of 8 weeks. 20 urine samples per subject and 1 serum sample per subject were collected before and during the probiotic assumpion. Univariate and multivariate statistical analyses were used to evaluate the 1H-NMR urine and INTRODUCTION serum spectra acquired, and to characterize the individual effects of the treatment. After the treatment, probiotics SPONSORS influence the urinary metabolic profiles of the volunteers, without altering their subject-specific phenotypes. In PROGRAM particular we observed a cumulative effect of probiotics during the time of the administration. Modifications in PROGRAM — BY DAY metabolite levels, especially in glucose, isoleucine, valine, 3-hydroxyisobutyricacid, 4-hydroxyphenylacetate, and TOPICS acetoaceticacid levels, were monitored. We observed, also, that probiotics influence the serum metabolic profiles, PRIZE LECTURES in particular they induce fluctuations in acetone, ascorbate, and citrate levels. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] Sommer, F. & Bäckhed, F. Nat. Rev. Microbiol. 2013, 11, 227–238. [2] Collins, M. D. & Gibson, G. R. Am. J. Clin. Nutr. 1999, 69, 1052S-1057S. [3] ADVERTISERS Martin, F.-P. J. et al. Mol. Syst. Biol. 2008, 4, 157. [4] Vignoli, A. et al. Angew. Chem. Int. Ed. 2019, 58, 968–994. [5]Ghini, V. et al. Metabolites 10, (2020). AUTHOR INDEX Acknowledgements: The authors acknowledge the Probiotical S.p.A (Novara) and specifically the CERM/CIRMMP Italy Centre. PO073 POSTERS / METABOLOMICS 226 DANIELA DUARTE1, BEATRIZ CASTRO1, SALIVA NMR METABOLOMICS IN THE SEARCH FOR JOANA LEONOR PEREIRA2, GESTATIONAL DIABETES MELLITUS BIOMARKERS JOANA FARIA MARQUES3, CRISTINA PITA4, FÁTIMA NEGRÃO4, MARIA DO CÉU ALMEIDA4, Gestational diabetes mellitus (GDM) is a carbohydrate intolerance with onset or first recognition during pregnancy ISABEL CARREIRA5, ANA LUÍSA COSTA2, [1]. Although diagnostic methods and therapies are available already, methods for the prediction of the condition ANA M. GIL1 and individual therapy response are still lacking. Hence, metabolomics of maternal urine and blood has searched 1CICECO-Aveiro Institute of Materials, Chemistry Department, for new GDM biomarkers [2] but little is known about the usefulness of saliva, a non-invasive and easy to collect University of Aveiro, Aveiro, Portugal 2 biofluid, which has emerged more recently as a valuable resource for disease research [3]. Dentistry Department, Faculty of Medicine, Institute of Paediatric and Preventive Dentistry, University of Coimbra, Coimbra, Portugal 3 This work reports a 1H NMR metabolomics study of saliva to characterize its stability under typical handling/ana-GIBBO-Oral Biology and Biochemistry Research Group, CEMBDE- COCHRANE Portugal-Faculty of Dental Medicine, University of Lisbon, lytical conditions of sampling in a clinical setting. Results show that saliva is stable up to 6h at room temperature Lisbon, Portugal and at 4ºC. Applied to pregnant women, the effects of different GDM treatments (diet, insulin and metformin) were 4Maternidade Bissaya Barreto, Centro Hospitalar e Universitário de Coimbra, Coimbra, Portugal explored through the saliva metabolome of the subjects. Multi- and univariate analysis suggested that the saliva 5Cytogenetics and Genomics Laboratory, and CIMAGO-Center for metabolic profile of GDM patients stabilizes upon diet treatment, while insulin or metformin therapies induce sev-Research in Environment, Genetics and Oncobiology, Faculty of eral more enduring changes, reflecting mostly the impact of diet and/or oral bacterial metabolism. The potential Medicine, University of Coimbra, Coimbra, Portugal and challenges of the use of saliva as a mean to aid in GDM diagnosis and management are discussed. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES References: [1] E B. E. Metzger, S. G. Gabbe, B. Persson, T. A. Buchanan, P. A. Catalano, P. Damm, A. R. Dyer, A. Leiva, M. Hod, J. L. Kitzmiler, L. P. Lowe, H.D. TUTORIAL LECTURE McIntyre, J. J. Oats, Y. Omori, M. I. Schmidt, International Association of Diabetes and Pregnancy Study Groups Consensus Panel, Diabetes Care. 2010 33, 676-682. [2] Q. Chen, E. Francis, G. Hu, L. Chen, J. Diabetes Complications. 2018, 32, 512-523. [3] A. Gardner, G. H. Carpenter, P. -W. So, Metabolites. 2020, 10, 47. INVITED AND PROMOTED LECTURES Acknowledgements: This project was developed within the scope of the projects CICECO-Aveiro Institute of Materials (UIDB/50011/2020 & UIDP/50011/2020), POSTERS financed by national funds through the Portuguese Foundation for Science and Technology (FCT)/ MEC and when appropriate cofinanced by FEDER under ADVERTISERS the PT2020 Partnership Agreement. We also acknowledge the Portuguese National NMR Network (RNRMN), supported by FCT funds and also FCT for the AUTHOR INDEX SFRH/BD/119509/2016 grant (DD). PO074 POSTERS / METABOLOMICS 227 ŠTĚPÁN HORNÍK1,2, NMR METABOLOMICS IN TOXICOLOGY: LENKA MICHÁLKOVÁ1,2, EFFECT OF NANOPARTICLES INHALATION ON BLOOD JAN SÝKORA1, VLADIMÍR ŽDÍMAL1, PLASMA AND EXHALED BREATH CONDENSATE DANIELA PELCLOVÁ3 PROFILES 1Institute of Chemical Process Fundamentals of the CAS, Prague, Although the application of nanoparticles (NPs) in different fields (electronics, optoelectronics, drug delivery and Czech Republic 2Department of Analytical medical diagnostics) has been growing extremely, concerns about adverse effects on public health, consumer safe-Chemistry, University of Chemistry ty and occupational safety are not properly described and understood [1]. Since NPs are able to cross cell barriers, and Technology, Prague, Czech enter cells and interact with subcellular structures, a common response to NPs exposure is the induction of oxi-Republic 3Department of Occupational dative stress and inflammation [2]. Potential risks of NPs exposure need to be assessed and biomarkers need to be Medicine, First Faculty of Medicine, identified to enable early diagnosis and prevention of occupational diseases. Surprisingly, the standard operating Charles University and General University Hospital, Prague, Czech procedures for monitoring NPs exposure at the work place are still insufficient. Republic Metabolomics offers a suitable strategy for analysis of individual subjects before and after NPs exposition, which allows to study perturbations in levels of metabolites and consequent alterations in metabolic pathways induced by acute toxicological effect. Moreover, comparison of subjects under long-term NPs exposition with healthy controls enables to assess chronic (subacute) toxicological effect [3]. In our study, 1H NMR metabolomics was used to analyse samples of exhaled breath condensate and blood plasma of pre- and post-shift researchers perfimng various tasks related to the processing of nanocomposite materials and healthy individuals, who do not encounter NPs in their INTRODUCTION occupation. The main aim of this work is to evaluate the effect of NPs inhalation. The altered metabolite levels were SPONSORS identified in both biofluids. Thus, the metabolic pathways affected by NPs exposure were determined and differenc- PROGRAM es in acute and chronic effects were observed. The achieved results confirm the applicability of NMR metabolomics PROGRAM — BY DAY in toxicological studies in the evaluation of the extent of toxic insult and understanding the molecular mechanism TOPICS of nanoparticle-organism interaction. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] S.H. Liou, C.S. Tsai, D. Pelclova, M.K. Schubauer-Berigan, P.A. Schulte, J. Nanopart. Res. 2015, 17, 413. [2] G. Canu, P.A. Schulte, M. Riediker, et ADVERTISERS al., J. Epidemiol. Community Health. 2018, 72, 148. [3] L.K. Schnackenberg, J. Sun, R.D. Beger, Methods Mol. Biol. 2012, 926, 141. AUTHOR INDEX Acknowledgements: The work was supported by the Czech science foundation (grant No. 18-020795). PO075 POSTERS / METABOLOMICS 228 KLEMEN PEČNIK1, IDENTIFICATION OF METABOLIC PROFILES SPECIFIC VESNA TODOROVIĆ2, FOR NORMAL AND TUMOR CELL LINES WITH THE MAŠA BOŠNJAK2, MAJA ČEMAŽAR2, GENERAL EXPLANATION METHOD AND NMR IGOR KONONENKO3, SPECTROSCOPY GREGOR SERŠA2, AND JANEZ PLAVEC1,4,5 Potential of machine learning is recognized rapidly in the field of systems biology, where valuable knowledge is hid-1National Institute of Chemistry, Slovenian den in large amounts of data produced by various analytical techniques including NMR. Machine learning models NMR Centre, Ljubljana, Slovenia 2 in metabolomics, despite their excellent prediction accuracy are still not adopted widely due to the lack of efficient Institute of Oncology, Department of Experimental Oncology, Ljubljana, Slovenia explanation of their predictions. In ourstudy we tested the use of the general explanation method [1] that enables 3University of Ljubljana, Faculty of Computer intuitive interpretation of arbitrary machine learning model’s predictions. The method was tested on a dataset of and Information Science, Ljubljana, Slovenia 4 1D 1H NMR spectra acquired on normal and tumor cell lines. The random forests and artificial neural network mod-EN-FIST Centre of Excellence, Ljubljana, Slovenia els were applied to a dataset and showed excellent prediction accuracy. Our results demonstrate that by using the 5University of Ljubljana, Faculty of general explanation method to explain models’ predictions, a concentration profiles of metabolites can be identi-Chemistry and Chemical Technology, Ljubljana, Slovenia fied that discriminate individual normal and tumor cell line types [2]. We believe that our results will encourage scientists to use the general explanation method and give rise to further in-depth exploration toward metabolic phenotyping, biomarker identification and therapeutic target detection of complex diseases through employment of machine learning in datasets that include clinical and biological information. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] E. Štrumbelj, I. Kononenko, J. Mach. Learn. Res. 2010, 11, 1–18. [2] K. Pečnik, V. Todorović, M. Bošnjak, M. Čemažar, I. Kononenko, G. Serša and AUTHOR INDEX J. Plavec, ChemBioChem. 2018, 19 (19), 2066-2071. PO076 POSTERS / METABOLOMICS 229 DESSISLAVA GERGINOVA, WHAT SPINS DIFFERENTLY IN PURE HONEY? SVETLANA SIMOVA The variety of new honey types connected with the exploitation of the plant biodiversity, the arising environmental Institute of Organic Chemistry with Centre of Phytochemistry, BulNMR, Sofia, and ecological issues as well as the traditional question for food authentication constantly require introduction of Bulgaria appropriate methods of analysis. Numerous challenges arise in trying to keep the natural and administrative regulations for ensuring the consumer protection. The content, the labeling and even the nature of many components within honey are subject of growing interest and investigations. In recent years a number of scandals in different counties show the utility of NMR spectroscopy as an important method for complete analysis, covering numerous aspects related to authenticity, quality control and quantification. Here, we present several recent investigations using the approuch of combined 13C NMR profiling combined with INTRODUCTION different chemometric approaches to differentiate botanical and geographical origin of honey, to find biomarkers SPONSORS and to detect adulteration on the example of honeys from the region (Bulgaria, North Macedonia) and from sting- PROGRAM less bees (Tanzania) [1,2,3]. PROGRAM — BY DAY The importance of honey as an effective natural therapy to attenuate acute inflammation is discussed in a recent TOPICS study describing the potential therapeutic effects of honey in the context of the COVID-19 pandemic [4]. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] D. Gerginova, Y. Mitrev, S. Simova, J. BAS, in press. [2] D. Gerginova, S. Simova, M. Popova, M. Stefova, J.Stanoeva, V. Bankova, Molecules 2020, INVITED AND PROMOTED LECTURES 25, 4687. [3] M. Popova, D. Gerginova, B. Trusheva, S. Simova, A. Tamfu, O. Ceylan, K. Clark, V. Bankova, Foods 2021, 10, 997. [4] K. Hossain, M. Hossain, A. POSTERS Moni, M. Rahman, M. U. Rahman, M. Alam, S. Kundu, M. Rahman, M. Hannan, M. Uddin, Heliyon 2020, 6 (12), e05798. ADVERTISERS Acknowledgements: The support by the Bulgarian Ministry of Education and Science under the National Research Program “Healthy Foods for a Strong Bio- AUTHOR INDEX Economy and Quality of Life” using the research equipment of the Distributed Research Infrastructure INFRAMAT is gratefully acknowledged. PO077 POSTERS / METABOLOMICS 230 MONIKA ŠKRJANC1, 2, NMR STUDIES OF SLOVENIAN HONEY DAMJAN MAKUC1, JANEZ PLAVEC1,2,3 Beekeeping is an important agricultural activity in Slovenia with a long and rich tradition. Slovenians are a bee-1 keeping nation as more than 10,000 inhabitants are engaged in this activity. Honey is an important and widely National Institute of Chemistry, Slovenian NMR Centre, Ljubljana, used food product for nutrient as well as in cosmetic and medicinal purposes. It is one of the most adulterated foods Slovenia worldwide[1] so the analysis and determination of chemical composition, especially carbohydrate composition, of 2University of Ljubljana, Faculty Slovenian honey are needed to protect this valuable product. NMR spectroscopy is powerful tool in food analysis as of Chemistry and Chemical Technology, Slovenia it provides important structural and chemical characterization. Honey is mainly composed of a rich repertoire of 3EN-FIST Centre of Excellence, sugars, fructose and glucose being predominant. The composition of sugars in honey depends on its geographical, Ljubljana, Slovenia botanical origin and other factors. Identification of carbohydrates is due to multiple isomeric forms and structural similarity very difficult. In our preliminary study we examined 25 samples of different types of Slovenian honey (forest, fir, linden, buckwheat, chestnut and acacia) with NMR in order to characterize chemical profiles specific for each group. Using 2D NMR homonuclear and heteronuclear techniques we achived assignment of carbohydrate peaks in 1D 1H spectrum of honey samples.[2] With PCA analysis we established key features for each group. Melezi-tose is the main metabolite found in forest and fir honey. Higher amounts of glucose can be found in buckwheat and linden honey while higher amounts of fructose are specific for chestnut and acacia honey. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] X. Zhou, M. P. Taylor, H. Salouros, S. Prasad, Sci. Rep. 2018, 8, 1–11. [2] G. D. Brown, J. Bauer, H. M. I. Osborn, R. Kuemmerle, ACS Omega 2018, AUTHOR INDEX 3, 17957–17975. PO078 POSTERS / METABOLOMICS 231 ALESSIA VIGNOLI1,2, CELIAC DISEASE AND NON-CELIAC GLUTEN SENSITIVITY LEONARDO TENORI1,2, TWO DIFFERENT GLUTEN-RELATED DISORDERS: ANTONIO CALABRÒ3, CLAUDIO LUCHINAT1,2 A SERUM METABOLOMICS AND LIPIDOMICS STUDY 1Magnetic Resonance Center (CERM) and Department of Celiac disease (CD) is an immune-mediated enteropathy triggered disease. Metabolomics has already proved to be a valuable instru-Chemistry “Ugo Schiff”, University of by the ingestion of gluten in genetically susceptible subjects. Un- ment for the study of the systemic metabolic alterations induced by Florence, Sesto Fiorentino, Italy 2 der the inclusive definition of celiac disease, potential celiac disease CD with respect to controls4–7. The present study aims at providing Consorzio Interuniversitario Risonanze Magnetiche (pCD) is defined by the presence of positive serum antibodies, HLA- further insights into the metabolic processes underpinning these MetalloProteine (CIRMMP), Sesto DQ2/DQ8 haplotypes, and a normal small intestinal mucosa (Marsh three gluten-related disorders (overt CD, pCD, GS) via NMR-based Fiorentino, Italy 3Department of Experimental grade 0-1)1. This condition occurs in one-fifth of CD patients and serum metabolomics and lipidomics. and Clinical Biomedical Sciences, usually represents a clinical challenge. In the setting of gluten-re- University of Florence, Italy Multivariate analysis on NMR data allowed the selective discrimi- lated disorders is also present the non-celiac gluten sensitivity nation between CD (overt and potential) and GS with high discrim- (GS), which has been defined as the development of intestinal and/ ination accuracies. In these discriminations both low molecular or extraintestinal symptoms related to the ingestion of gluten in weight metabolites as well as lipoproteins significantly contribute. patients without celiac disease2. In GS neither allergic nor autoim- The most relevant pathways identified are Glyoxylate and dicarbox- mune mechanisms are involved, and is estimated that affected 6% ylate metabolism and Glycolysis/ Gluconeogenesis. Whereas oCD of the European population3. This disease and its pathogenesis are and pCD show only slight differences (accuracy ~65%). yet poorly understood and relationships between GS and CD need to be further investigated. There are no diagnostic biomarkers avail- In conclusion, NMR-based metabolomics seems to be effective in INTRODUCTION able for GS, therefore the diagnosis is complex as symptoms overlap discriminating gluten-related disorders, this is particularly relavant SPONSORS with those of CD, irritable bowel syndrome and inflammatory bowel for GS because no diagnostic tools are currently available. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] C. M. Trovato, et al., Gastroenterol Res Pract 2019, DOI 10.1155/2019/8974751. [2] A. Sapone, et al., BMC Med 2012, 10, 13. [3] C. Catassi, et al., Nutrients 2013, 5, 3839–3853. POSTERS [4] I. Bertini, A. Calabrò, V. De Carli, C. Luchinat, S. Nepi, B. Porfirio, D. Renzi, E. Saccenti, L. Tenori, J. Proteome Res. 2008, 8, 170–177. [5] P. Bernini, I. Bertini, A. Calabrò, G. la Marca, G. Lami, ADVERTISERS C. Luchinat, D. Renzi, L. Tenori, J. Proteome Res. 2010, 10, 714–721. [6] A. Calabrò, Antonio, E. Gralka, C. Luchinat, E. Saccenti, L. Tenori, Autoimmune Diseases 2014, 2014, e756138. [7] A. AUTHOR INDEX Vignoli, B. Orlandini, L. Tenori, M. R. Biagini, S. Milani, D. Renzi, C. Luchinat, A. S. Calabrò, J. Proteome Res. 2019, 18, 1228–1236. PO079 POSTERS / HYPERPOLARIZATION 232 KERTI AUSMEES, TOWARDS PH HYPERPOLARIZED ANALYSIS OF URINE 2 NELE REIMETS, WITH MINIMAL SAMPLE MANIPULATION INDREK REILE National Institute of Chemical Urine is an easily obtainable highly complex biofluid containing thousands of compounds, with various chemical/ Physics and Biophysics, Tallinn, physical properties and concentrations [1]. Ability to analyze these compounds is of great diagnostic and scientific Estonia interest. Nuclear magnetic resonance (NMR) is one of the principle analytical techniques used for the purpose, but many urinary metabolites occur below the NMR limit of detection (LOD). Sensitivity can be improved by Iridium catalyzed pH hyperpolarized chemosensing (pH -HP) [2], where additional spectral resolution can be derived from 2 2 2D zero-quantum (ZQ) spectroscopy [3]. This method can detect nanomolar concentrations of nitrogen containing compounds [3]. However, direct analy- sis of urine is challenging, because of high quantity of ammonia in urine. Previously solid phase extraction (SPE) methods have been used to remove ammonia and other pH -HP interfering components like water, salts, protein 2 and urea from urine. The SPE organic solvent provides optimal working conditions for the iridium catalyst [4]. The drawback of SPE based methods is the tendency to decrease overall metabolite coverage and, hence, SPE is often avoided in global analysis of compound in biofluids [5]. Herein we present the results of our current work, where ammonia, urea and water are removed from urine sample by raising the pH and freeze drying. Resulting solids are reconstituted with water and salts are precipitated with methanol. The resulting methanol-water mixture is compatible with the usual Iridium based pH -HP chemosens-2 INTRODUCTION ing catalyst and results in a reproducible hyperpolarized spectrum allowing to detect several endogenous metabo- SPONSORS lites that occur below the LOD of regular NMR. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] S. Bouatra, F. Aziat, R. Mandal, A. C. Guo, M. R. Wilson, C. Knox, T. C. Bjorndahl, R. Krishnamurthy, F. Saleem, P. Liu, Z. T. Dame, J. Poelzer, J. Huynh, F. S. Yallou, N. Psychogios, E. Dong, R. Bogumil, C. Roehring, D. S. Wishart, PLoS One 2013, 8, e73076. [2] N. K. J. Hermkens, N. Eshuis, B. J. A. van POSTERS Weerdenburg, M. C. Feiters, F. P. J. T. Rutjes, S. S. Wijmenga, M. Tessari Anal. Chem. 88, 3406–3412, 2016 [3] L. Sellies, I. Reile, R. L. E. G. Aspers, M. C. Feiters, ADVERTISERS and F. P. J. T. Rutjes, M. Tessari Chem. Commun. 2019, 55, 7235–7238, [4] N. Reimets, K. Ausmees, S. Vija, I. Reile Anal. Chem. , under review [5] 1. Sitnikov, D. G., AUTHOR INDEX Monnin, C. S. & Vuckovic, D. Sci. Rep. 2016, 6, 38885. PO080 POSTERS / HYPERPOLARIZATION 233 SAUMYA BADONI1, PROBING LIGAND COORDINATION DRIVEN MICHAŁ TERLECKI2,3, SHAPE-SELECTIVE GROWTH OF NANOPARTICLES BY MAŁGORZATA WOLSKA- PIETKIEWICZ2, DNP NMR SPECTROSCOPY DANIEL LEE1, JANUSZ LEWIŃSKI2,3, Zinc oxide nanoparticles (NPs) find applications in numerous fields such as solar cells, sensors, medicine, paint, and GAËL DE PAËPE1 rubber industries [1]. This has motivated researchers to optimize synthetic procedures to produce NPs with desired 1 applications. The shape, such as spherical, rod-like or platelet, is among the key features that can be modulated Université Grenoble Alpes, CEA, INAC-MEM, Grenoble, France to obtain specific function. Thus, there is a need to investigate the interface of these materials in order to control 2Faculty of Chemistry, Warsaw their morphology which is largely dependent on the nanocrystal-ligand interactions at this interface [2]. Among University of Technology, Warsaw, contemporary analytic techniques, DNP-enhanced solid-state NMR spectroscopy is highly suitable for character- Poland 3Institute of Physical Chemistry, Polish izing these interfaces due to its site-specific detection ability coupled with significant gain in NMR sensitivity. In Academy of Sciences, Warsaw, Poland this presentation, it will be discussed how MAS-DNP NMR in combination with complementary techniques such as density functional theory (DFT) calculations and HRTEM, can be used to determine ligand coordination modes and atomic-scale arrangements on bi-faceted hexagonal ZnO nanoplatelets, that are typically used to produce high-ly-ordered ultra-thin (< 5nm) films which are considered as prime candidates for new generation optoelectronic devices [3]. Moreover, insights into the growth of these NPs will be provided. This work relating ligand binding modes and arrangement with particle morphology will allow controlled production of rationally-designed nanocrystals with specific functions. INTRODUCTION More specifically, DNP enhanced 13C homonuclear dipolar correlation spectra for carbon peaks from surface ligands SPONSORS at 13C natural isotopic abundance will be presented. These were used to study ligand proximity and arrangement of PROGRAM ligands on the ZnO surface. Also, DNP-enhanced 13C and 15N MAS NMR spectra and 1H-15N heteronuclear correla- PROGRAM — BY DAY tions, recorded at natural isotopic abundance will be discussed. These spectra evidence the presence of various TOPICS ligand-binding sites at the inorganic interface and these will be associated with different facets of the ZnO nano- PRIZE LECTURES platelets. The rich structural information obtained from the NMR experiments is complemented with DFT chem- PLENARY LECTURES ical shift calculations of various ZnO surface models to help provide a full picture of the nanoplatelet interfaces. TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Moezzi, A.; McDonagh, A. M.; Cortie, M. B. Chem. Eng. Sci. 2012, 185, 1-22. [2] Bealing, C. R.; Baumgardner, W. J.; Choi, J. J.; Hanrath, T.; Hennig, AUTHOR INDEX R. G. ACS nano 2012, 6, 2118-2127. [3] Nasilowski, M., Mahler, B., Lhuillier, E., Ithurria, S. and Dubertret, B. Chemical reviews 2016, 116(18), 10934-10982. PO081 POSTERS / HYPERPOLARIZATION 234 THOMAS BIEDENBÄNDER, THE ROLE OF METHYL DYNAMICS IN DNP VICTORIA ALADIN, BJÖRN CORZILIUS Methyl NMR studies have become very popular during the last de- dynamics/crowding [4]. However, until now, no studies of methyl cades. The fast three-fold reorientation of the methyl group around dynamics particularly under DNP conditions were performed. Thus, Instiute of Chemsitry and Department of Life, Light and Matter, University of its symmetry axis (H C-C) yields advantageous relaxation properties the aim of this project is the measurement of the three-fold reori- 3 Rostock, and thus well-resolved NMR spectra [1]. For instance, these proper- entation dynamic under DNP conditions in various methyl-bearing Rostock, Germany ties are utilized in methyl TROSY HMQC which is commonly used molecules. Therefore, we studied three different methyl-bearing for structure determination of large biomolecules. Moreover, it was amino acids in which the methyl protons were selectively deuter-recently shown that the three-fold reorientation is still active under ated in polycrystalline powder. The selective deuteration enabled DNP conditions [2]. The observed effect is exploited in SCREAM- us to determine the correlation time of the three-fold reorientation DNP (Specific Cross Relaxation Enhancement by Active Motions dynamics by T relaxation measurement in polycrystalline samples. 1 under DNP). Here, polarization transfer from the hyperpolarized The activation energy of the reorientation was further calculated by 1H spins to 13C is driven by the cross-relaxation-promoting methyl applying the Arrhenius equation yielding activation energies be- dynamics. The specifically hyperpolarized methyl-13C can then be tween 8 and 21 kJ/mol. The calculated activation energies show a used, for example, for either measuring the binding of a ligand to an large difference for Ala-C D and Met-C D , which explains the low- β 3 ε 3 RNA aptamer [3] or to membrane proteins such as the green absorb- er observed enhancement factor for Ala due to the higher energy ing proteorhodopsin [4]. barrier for the cross relaxation-promoting three-fold reorientation. For a more detailed molecular understanding of the mechanism, the Experiments under DNP conditions of selectively deuterated mol- enhancement factors of SCREAM-DNP of all methyl-bearing amino ecules are currently underway. The preliminary results suggest a INTRODUCTION acids were determined in a temperature range from 110 K to 165 distribution of T relaxation rates represented by a stretched expo-1 SPONSORS K [5]. Therein, a clear correlation between the DNP enhancement nential function as it was also shown in wetted protein powder [6]. PROGRAM and the expected methyl dynamics has been shown, providing the However, these experiments have to be conducted in more detail. PROGRAM — BY DAY prospect of using SCREAM-DNP as a quantitative measure for local TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Wiesner, S. & Sprangers, R. Curr. Opin. Struct. Biol. 2015, 35, 60–67. [2] Daube, D., Aladin, V., Heiliger, J., Wittmann, J. J., Barthelmes, D., Bengs, C., Schwalbe, H., & Corzilius, B. POSTERS J. Am. Chem. Soc. , 2016, 138, 16572–16575. [3] Aladin, V., Vogel, M., Binder, R., Burghardt, I., Suess, B., & Corzilius, B. Angew. Chem. Int. Ed. 2019, 131, 4917–4922. [4] Mao, J., Aladin, V., Jin, X., ADVERTISERS Leeder, A. J., Brown, L. J., Brown, R. C. D., He, X., Corzilius, B., & Glaubitz, C. J. Am. Chem. Soc. 2019., 141(50), 19888–19901. [5] Aladin, V. & Corzilius, B. Solid State Nucl. Magn. Reson. 2019, 99, AUTHOR INDEX 27–35. [6] Vugmeyster, L., Ostrovsky, D., Penland, K., Hoatson, G. L., & Vold, R. L. J. Phys. Chem. B 2013, 117, 1051–1061. PO082 POSTERS / HYPERPOLARIZATION 235 VASYL DENYSENKOV, TRIPLE RESONANCE (E, 1H, 13C) PROBEHEAD FOR DANHUA DAI, LIQUID-STATE DNP EXPERIMENTS AT 9.4 TESLA THOMAS F. PRISNER Institute for Physical and In DNP experiments, nuclear spin polarization is enhanced by transferring the relatively larger electron polariza-Theoretical Chemistry, Goethe tion to target nuclei via microwave irradiation. Here, we describe the design and performance of a probehead for University Frankfurt, and Center of Biomolecular Magnetic Resonance, Overhauser DNP experiments on protons and 13C in liquid samples of about 100 nanoliters. Proton decoupling un- Frankfurt am Main, Germany der DNP conditions as a new tool is possible with the developed probehead. Besides, heat dissipation was improved with respect to the DNP probeheads described before [1, 2] that helped to keep aqueous samples at constant temperature under irradiation by microwave power up to 5 Watts. This feature opens opportunity to disentangle tem- perature and microwave saturation effects on DNP enhancement in liquids. Performance of the probe was tested by DNP experiments on aqueous 3-13C-sodium pyruvate solutions. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] V. Denysenkov, T.F. Prisner, J. Magn. Reson. 2012, 217, 1-5. [2] V.P. Denysenkov, M.J. Prandolini, A. Krahn, M. Gafurov, B. Endeward, and T.F. AUTHOR INDEX Prisner, Appl. Magn. Reson. 2008, 34, 289-299. PO083 POSTERS / HYPERPOLARIZATION 236 STUART J. ELLIOTT1,2, A 3D-PRINTED 1H-13C BACKGROUND-FREE MORGAN CEILLIER1, RADIOFREQUENCY COIL ENABLES SIMPLE OLIVIER CALA1, QUENTIN STERN1, IMPLEMENTATION OF CROSS-POLARIZATION AT SAMUEL F. COUSIN1,3, LOW TEMPERATURE UNDER DISSOLUTION-DYNAMIC THÉO EL DARAÏ1 AND SAMI JANNIN1 NUCLEAR POLARIZATION CONDITIONS 1Centre de Résonance Magnétique Nucléaire à Très Hauts Champs - The low sensitivity of conventional nuclear magnetic resonance experiments can be readily overcome by employ- Université de Lyon / CNRS / Université ing hyperpolarization methodologies, in suitable cases. One such technique, dissolution-dynamic nuclear po- Claude Bernard Lyon 1 / ENS de Lyon, larization ( d DNP), provides a robust means of strongly polarizing a variety of small molecules. A drawback of the Villeurbanne, France 2Department of Chemistry, University d DNP approach, the excessively long polarization timescales for insensitive nuclei, has been circumvented using of Liverpool, Liverpool, UK cross-polarization (CP) pulse sequences [1,2], which are in general quicker and more efficient (>10× faster polariza-3Institut de Chimie Radicalaire - UMR 7273, Saint-Jérôme Campus, Av. Esc. tion of 13C nuclei for d DNP, typically improving from ~2 hours to ~10 minutes). However, the capacity to effectively Normandie Niemen, Aix-Marseille perform CP experiments under d DNP conditions remains challenging and is still plagued by additional complica-Université / CNRS, Marseille, France tions including spurious background signals from the experimental apparatus. Here we propose a background-free 1H-13C radiofrequency coil specifically designed for use in CP experiments at liquid helium temperatures, which allows simplified and “on-the-spot” nuclear spin polarization quantification. We additionally introduce simple guidelines for the optimization and implementation of CP pulse sequences. The background-free 1H-13C radiofrequency coil is straightforward to construct [3], fabricated from 3D printed copper/silver wire and ceramic capacitors, and is INTRODUCTION housed in a rigid PTFE support. Experimental demonstrations are presented for the case of [1-13C]sodium acetate. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] A. Bornet, R. Melzi, S. Jannin and G. Bodenhausen, Appl. Magn. Reson. 2012, 43, 107-117. [2] A. Bornet, A. Pinon, A. Jhajharia, M. Baudin, X. Ji, L. Emsley, G. Bodenhausen, J.-H. Ardenkjær-Larsen and S. Jannin, Phys. Chem. Chem. Phys. 2016, 18, 30530-30535. [3] A. Bornet, R. Melzi, A. J. Perez Linde, P. INVITED AND PROMOTED LECTURES Hautle, B. van den Brandt, S. Jannin and G. Bodenhausen, J. Chem. Phys. Lett. 2013, 4, 111-114. POSTERS Acknowledgements: This research was supported by ENS-Lyon, the French CNRS, Lyon 1 University, Bruker Biospin, the European Research Council under ADVERTISERS the European Union’s Horizon 2020 research and innovation program (ERC Grant Agreements No. 714519 / HP4all and Marie Skłodowska-Curie Grant AUTHOR INDEX Agreement No. 766402 / ZULF). PO084 POSTERS / HYPERPOLARIZATION 237 M. VICTORIA GÓMEZ1, PHOTO-CIDNP NMR AS A TOOL TO INVESTIGATE MARGARITA RUIZ- CONFORMATIONAL TRANSITIONS IN SWITCH PEPTIDES CASTAÑEDA1, PHILIPP NITSCHKE2, Photo-Chemically Induced Dynamic Nuclear Polarization (Pho- Herein, we report valuable residue-level information by Photo-CIDNP RUTH M. GSCHWIND2, to-CIDNP) is an excellent probe to scrutinize the solvent exposure experiments that affords a clear picture of the conformational transi-M. ANGELES JIMÉNEZ3 of individual tryptophan, tyrosine or histidine amino acids in a se- tion of LytA239-252 peptide from the native β-hairpin into the a-helix trig- 1IRICA, Faculty of Chemical Sciences quence, i.e. which of those aromatic side chains are participating gered by the presence of DPC micelles. We show that aromatic stacking and Technologies, Universidad de in an interaction (hindered from the solvent) and which are freely and disruption encodes the key for the conformational switching. Fur-Castilla-La Mancha (UCLM), Ciudad Real, Spain exposed. This approach involves a spin-selective photochemical thermore, considering that when executing Photo-CIDNP experiments, 2Institute of Organic Chemistry, process between the target molecule and a laser-excited photosen- the uniform illumination of the whole sample volume faces several University of Regensburg, sitizer dye and can lead to positive and negative enhancements of problems, we describe the whole optimization process for the NMR ex-Regensburg, Germany 3Departamento de Química-Física NMR signals. Interestingly, signal enhancements produced by Pho- perimental conditions, illustrating the advantages of using low NMR Biológica, Instituto de Química Física to-CIDNP can be transferred between nearby nuclei in a mechanism active volumes to gain all information from the spectra. In this sense, Rocasolano, Madrid similar to the NOE effect, namely cross polarization, which can be our previous experience [3] in the development of in-situ NMR illumi-interpreted to give an idea about the mobility of a molecule [1]. nation devices for pushing the NMR sensitivity for small sample vol- umes, supports the importance of the optimization of the photon flux These interesting features encouraged us to use photo-CIDNP to gain in the NMR active volume. insights into the micelle-triggered conformational transition observed in LytA239-252, a surface protein fragment present in pneumococcus, the Our results herein, demonstrate the great potential of using Pho- most common pathogen of the respiratory tract that causes pneumo- to-CIDNP experimental data in an optimized set of experimen- INTRODUCTION nia. LyTA239-252 is a fragment of a choline-biding protein that are essen- tal conditions to determine conformational transitions of switch SPONSORS tial for bacterial colonization and virulence, with the involvement of peptides. PROGRAM conformation transition [2]. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] L. Zetta, R. Kaptein, P. Hore, Febs Letters, 1982, 145, 277-280. [2] H. Zamora, B. Maestro, E. Strandberg, A. S. Ulrich, M. J. Sanz, M. A. Jimenez, Chemistry-A European Journal 2015, ADVERTISERS 21, 8076-8089. [3] M. Mompean, R. M. Sanchez-Donoso, A. Hoz, V. Saggiomo, A. H. Velders, M. V. Gomez, Nat. Commun. 2018, 9:108. AUTHOR INDEX Acknowledgements: MINECO is acknowledged for financial support (CTQ2017-84825-R; CTQ2017-84371P). PO085 POSTERS / HYPERPOLARIZATION 238 SANJAY VINOD KUMAR, NUMERICAL SIMULATIONS OF TRIPLET STATE GENERATION IN XIAOXUN CHEN, PHOTO-DNP SAMPLES GUINEVERE MATHIES Department of Chemistry, University In photo-DNP, light-excitation of a suitable molecular system is used to cre- N , N , N are the population densities of the S , S and T of Konstanz, Konstanz, Germany 0 1 T 0 1 1 ate a high, non-Boltzmann polarization, which is then transferred to nu- states, and N is the photon density. With the script, we cal- p clei to enhance the sensitivity of an NMR measurement. Perhaps the best- culated how the triplet states become populated, even after known example is triplet-DNP, introduced by Wenckebach et al. , in which the excitation source is switched off, and how the triplet the polarization of the excited triplet state of pentacene is transferred to population decays over time, depending on the phospho-1H in the naphthalene matrix by the integrated solid-effect.[1] To obtain an rescence lifetime (t = 1/(k +k )). It allowed us to optimize Ph Ph ICT optimized and uniform enhancement of the 1H spin polarization through- parameters such as laser pulse length and energy to gener- out the entire sample in these and other photo-DNP experiments, the exci- ate ~100% triplet state population, given the photophysical tation light source must be appropriately chosen and match with the con- properties such as fluorescence lifetime (t ) and ISC quan- F centration as well as the photo-physical properties of the photo-sensitive tum yield (F ). ISC molecules. To this end, numerical simulations are indispensible. We used the script to investigate the triplet state generation With the help of MATLAB, we wrote a script to simulate how the triplet in photo-DNP experiments with quinone-TEMPO molec-state population changes with time (t) and penetration depth (z) of a laser ular complexes in frozen toluene [4], which were recently beam into the sample. The script is based on the rate equations provided performed in our laboratory. In these experiments, we used by Takeda et al.,[2] given here in the notation of Wenckebach.[3] an excimer laser (pulse length of 5 ns, pulse energy 2 mJ) as the light source. We measured the photophysical properties INTRODUCTION for the quinones and quinones/TEMPO complexes in frozen SPONSORS …(1) toluene and methylcyclohexane to provide the script with PROGRAM realistic parameters. According to our calculations, the trip- PROGRAM — BY DAY …(2) let state population was only a few percent, explaining the TOPICS very modest enhancements we observed. Specifically, the PRIZE LECTURES …(3) calculations suggested that the photo-DNP efficiency in our PLENARY LECTURES experiments can be strongly improved by choosing a more TUTORIAL LECTURE …(4) suitable excitation source. INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] A. Henstra, T.S. Lin, J. Schmidt, W. Th. Wenckebach, Chem. Phys. Lett. 1990, 6-10. [2] K. Takeda, K. Takegoshi, T. Terao. J. Chem. Phys. 2002, 117, 4940-4946. [3] W.Th. Wenckebach, AUTHOR INDEX Dynamic Nuclear Polarisation using Photoexcited triplet states, 2008. [4] V. F. Tarasov, I. A. Shkrob, A. D. Trifunac. J. Phys. Chem. A. 2002, 106, 4838-4845. PO086 POSTERS / HYPERPOLARIZATION 239 MARCEL LEVIEN1,2, SPIN DENSITY ACCESSIBILITY AND LOCAL DYNAMICAL MAIK REINHARD1,2, FLUCTUATIONS OF ORGANIC RADICALS AFFECT LIQUID-STATE MARKUS HILLER1, IGOR TKACH1, DNP EFFICIENCY TOMAS ORLANDO1, Within the last decade, the polarization transfer mechanisms of density on the PA needs to be accessible and highly localized. In MARINA BENNATI1,2 Overhauser dynamic nuclear polarization (ODNP) in the liquid state particular, the solvent accessible surface (SAS), calculated for the 1 have been extensively studied for nuclei such as 1H and 13C. For large positions with highest spin density, correlates well with our results Max-Planck-Institute for Biophysical Chemistry, signal enhancements (102 – 103) at high magnetic fields, room tem- of the DNP efficiency. Indeed, a reduced accessibility of the radical Göttingen, Germany perature and ambient pressure, a scalar polarization transfer (Fer- site should change the likelihood of an encounter between PA and a 2University of Göttingen, mi contact interaction) is beneficial. Scalar polarization transfer is target, and therefore influence the frequency of such collisions [3]. Department of Chemistry, Göttingen, Germany mediated by molecular collision, whose frequency and duration are Additionally, we closely investigated the case of FN2a, whose high usually short enough (ps or sub-ps) to still contribute to the cross-re- signal enhancement at low magnetic fields (1.2 T) rapidly decays laxation rates at magnetic fields higher than 9.4 T. This is particular- at high fields (9.4 T) and there is outperformed by smaller NODs ly true for 13C nuclei [1, 2]. by almost a factor of 40 [2]. Field dependent DNP measurements To highlight how the scalar mechanisms critically depend on the suggested an additional contribution to the scalar relaxation with choice of the polarizing agent (PA)/ target molecule system, we a longer correlation time (~10 ps). This contribution is absent for performed a 13C ODNP study on six polarizing agents, five nitrox-smaller NODs. Complementary molecular dynamic simulations ide derivatives (NODs) and a,γ-bisdiphenylene-phenylallyl (BDPA) (MD) of FN2a show a rapid reorientation of the methyl groups on in model systems at 1.2 T. As a measure for the efficiency of each a timescale that correlates well with the one extracted from the INTRODUCTION PA, we use the coupling factor ξ extracted from the Overhauser field-dependent DNP data [4]. SPONSORS equation. We identified features that affect the DNP efficiency by In summary, we identified a field independent (a localized and acces- PROGRAM approximately a factor of five and observed a clear trend in DNP ef- sible spin density) as well as a field dependent (rapid local structural PROGRAM — BY DAY ficiency going from the least efficient PA (BDPA), over small NODs, fluctuations) feature of efficient PAs for DNP in the liquid state at low TOPICS and up to the most efficient FN2a (a nitroxide linked to a fullerene). and high magnetic field. Similar to solid-state DNP-NMR, where the PRIZE LECTURES Interestingly, this behavior is unique for the scalar mechanism, and new design of PAs is a thriving area for DNP optimization, our results PLENARY LECTURES has not been observed in comparative 1H-DNP measurements [3, 4]. should help to significantly boost ODNP in the liquid state. TUTORIAL LECTURE Supported by a DFT analysis, we recognized that the electron spin INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] G. Liu, et al. , Nat. Chem. 2017, 9, 676-680. [2] T. Orlando, et al. , Angew. Int. Ed. 2019, 58, 1402-1406. [3] M. Levien, et al. , Phys. Chem. Chem. Phys, 2021, 23, 4480-4485. [4] M. AUTHOR INDEX Levien, et al. , J. Phys. Chem. Lett. 2020, 11, 1629-1635. PO087 POSTERS / HYPERPOLARIZATION 240 ANDRIY MARKO, SIMULATION OF NITROGEN NUCLEAR SPIN ANTONIN SOJKA, MAGNETIZATION OF LIQUID SOLVED NITROXIDES OLEKSII LAGUTA, PETR NEUGEBAUER Nitroxide radicals are widely used in Electron Paramagnetic Resonance (EPR) applications. Nitroxides are stable Central European Institute of organic radicals containing N-O group with hyperfine coupled unpaired electron and nitrogen nuclear spins. In Technology - Brno University of the past, much attention was devoted to studying nitroxide EPR spectra and electron spin magnetization evolution Technology (CEITEC-BUT), Brno, Czech Republic under various experimental conditions. However, the dynamics of nitrogen nuclear spin was not investigated in detail so far. In this work, we performed quantitative prediction and simulation of nitrogen nuclear spin magnetization evolution in several magnetic resonance experiments. Our research was focused on fast rotating nitroxide radicals in liquid solutions. We used a general approach allowing us to compute electron and nitrogen nuclear spin magnetization from the same time-dependent spin density matrix obtained by solving the Liouville/von Neumann equation [1]. Especially we want to emphasize our prediction of a large dynamic nuclear polarization of nitrogen upon nitroxide irradiation with microwaves and influence of nitrogen nuclear spin dynamics on the nitroxide EPR saturation factor [2,3]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] Abragam, A., Principles of Nuclear Manetism, Oxford University Press, 2011. [2] Sezer, D., Gafurov, M., Prandolini, M. J., Denysenkov, V. P., & Prisner, T. F., Phys. Chem. Chem. Phys., 2009, 11, 6638. [3] Neugebauer, P., Krummenacker, J. G., Denysenkov, V. P., Parigi, G., Luchinat, C., & Prisner, T. F., Phys. INVITED AND PROMOTED LECTURES Chem. Chem. Phys., 2013, 15, 6049. POSTERS Acknowledgements: AM and OL are grateful for the support of their research work by the ESF under the project CZ.02.2.69/0.0/0.0/19\_074/0016239. PN ADVERTISERS acknowledges funding from the ERC under the European Union's Horizon 2020 research and innovation program (GA No. 714850). This work was also AUTHOR INDEX supported by Ministerstvo Školství, Mládeže a Tělovýchovy via CEITEC 2020 (LQ1601). PO088 POSTERS / HYPERPOLARIZATION 241 NELE REIMETS, DEVELOPING ANALYTICAL APPLICATIONS KERTI AUSMEES, FOR PARAHYDROGEN HYPERPOLARIZATION: SIRJE VIJA, INDREK REILE URINARY ELIMINATION PHARMACOKINETICS OF NICOTINE National Institute of Chemical Physics and Pharmacokinetics is essential in modern drug development in combining pH HP and 2D spectroscopy, it is possible to overcome 2 Biophysics, Tallinn, Estonia order to understand the (bio)chemical processes that a drug un- the NMR sensitivity barrier and detect midnanomolar concentra- dergoes in the body from its intake to excretion. To follow minute tions of a drug and a drug metabolite in a biofluid matrix [5]. concnetrations of analytes in complex mixtures like urine or blood, Herein, we demonstrate how pH HP can be applied in pharma- exceptional resolution and sensitivity is desired. Nuclear magnetic 2 cokinetics. As a proof of concept, we measured urine of humans resonance spectroscopy (NMR) is an attractive choice for PK for it exposed to nicotine and compared two common intake methods offers straightforward data interpretation and quantitative analysis. – smoking and absorption through skin by using a transdermal However, pharmacologically active compounds and their metabo- patch. We followed urinary elimination of nicotine, and its main lites in biofluids often appear in minute concentrations, well below metabolite cotinine, and determined their concentrations in urine the detection limit of NMR. during the onset and withdrawal from nicotine consumption. An To improve the NMR sensitivity parahydrogen (pH ) hyperpolar- NMR limit of detection of 0.1 μM and a limit of quantitation of 0.7 2 ization (HP) has been adapted for measuring biofluids [1]. Recent μM was achieved in a practical pharmacokinetics scenario where progress in pH HP techniques allows multiscan experiments [2], [3] precise quantitative and qualitative analysis is desired. 2 and to resolve complex spectra of urine with 2D spectroscopy [4]. By INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] I. Reile, N. Eshuis, N. K. J. Hermkens, B. J. A. van Weerdenburg, M. C. Feiters, F. P. J. T. Rutjes, M. Tessari, Analyst 2016, 141, 4001–4005. [2] N. Eshuis, R. L. E. G. Aspers, B. J. A. van Weerdenburg, M. C. Feiters, F. P. J. T. Rutjes, S. S. Wijmenga, M. Tessari, Angew. Chemie Int. Ed. 2015, 54, 14527–14530. [3] N. K. J. Hermkens, N. Eshuis, B. J. A. van Weerdenburg, M. C. Feiters, F. INVITED AND PROMOTED LECTURES P. J. T. Rutjes, S. S. Wijmenga, M. Tessari, Anal. Chem. 2016, 88, 3406–3412. [4] L. Sellies, I. Reile, R. L. E. G. Aspers, M. C. Feiters, F. P. J. T. Rutjes, M. Tessari, Chem. Commun. 2019, 55, 7235–7238. POSTERS [5] N. Reimets, K. Ausmees, S. Vija, I. Reile, Anal. Chem. ( under review 2021). ADVERTISERS Acknowledgements: This work was financially supported by the Estonian Research Council grant PSG11 and the Center of Excellence TK134 of the Archimedes Foundation. We thank Dr. Jaan AUTHOR INDEX Past for developing bubbling control system electronics. PO089 POSTERS / HYPERPOLARIZATION 242 QUENTIN STERN1, ELECTRON DRIVEN SPIN DIFFUSION UNDER DYNAMIC NUCLEAR ALESSANDRO CHESSARI1, POLARIZATION AT LOW TEMPERATURE SAMUEL F. COUSIN1, FRÉDÉRIC MENTINK-VIGIER2, Dynamic nuclear polarization (DNP) is a widely used tool for over- observing the flow of polarization between them. We have demon- ARTHUR C. PINON3, coming the low intrinsic sensitivity of magnetic resonance spec- strated the versatility of our approach in 1H DNP experiments with STUART J. ELLIOTT4, troscopy and imaging. DNP relies on transferring the high polariza- TEMPOL radicals conducted between 1.2 and 4.2 K in static mode OLIVIER CALA1 AND tion of unpaired electrons to nuclear spins via microwave irradia- and at 100 K under magic angle spinning (MAS) - conditions typi- SAMI JANNIN1 tion. Nuclei which are too far from the electron to interact directly cal for dissolution-DNP and MAS-DNP - and directly observed the 1CRMN- UMR 5280 Université de Lyon / CNRS with it may still be polarized indirectly by nuclear spin diffusion dramatic dependence of polarization flow on temperature. Using / Université Claude Bernard Lyon 1 / ENS de from the nuclei closer to the electron. However, the closest nuclear broadband adiabatic inversion pulses to manipulate the non-ob-Lyon, Villeurbanne, France 2National High Magnetic Field Laboratory, spins, even if they are potentially most efficiently polarized by the servable spins near the electron, we further showed that nuclear Florida State University, Tallahassee, FL, USA electron, have their Larmor frequencies so strongly shifted by the spins as close as 3 Å to the electron could still exchange polariza-3Swedish NMR Centre, University of interaction with the electron that they cannot exchange polariza- tion with bulk nuclei [3-4]. Gothenburg, Gothenburg, Sweden 4Department of Chemistry, University of tion with the bulk spins. These nuclei are said to be within the so- Liverpool, Liverpool, UK We now show similar findings for 13C DNP with trityl radicals be- called ‘spin diffusion barrier’ [1]. tween 1.2 and 4.2 K and investigate possible mechanisms for these A quantitative assessment of this barrier used to be hindered by the spin diffusion processes. We find that the fluctuation of the elec- lack of general methods for studying nuclear polarization flow in tron spin state could be responsible for compensating the energy the vicinity of paramagnetic centers. We have recently filled this mismatch between |aβ> and |βa> nuclear spin states and therefore gap by introducing a general set of experiments based on micro-allow nuclear spin diffusion [5]. These fluctuations are quenched INTRODUCTION wave gating that can be readily implemented [2]. The hyperpolar- when the electron polarization reaches 100%, which explains why SPONSORS ization resurgence experiments (HypRes) consist of generating a diffusion is slower at 1.2 K. PROGRAM large polarization gradient between the core and bulk spins and PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] W. E. Blumberg, Phys. Rev. 1960, 119(1), 79–84. [2] Q. Stern, S. F. Cousin, F. Mentink-Vigier, et al., Sci. Adv. 2021, 7 [3] J. P. Wolfe, Phys. Rev. Lett. 1973, 31(15), 907–910. [4] K. Ooi INVITED AND PROMOTED LECTURES Tan, M. Mardini, C. Yang et al., Sci. Adv. 2019, 5. [5] E. P. Horvitz, Phys. Rev. B. 1971, 3(9), 2868–2872. POSTERS Acknowledgements: This research was supported by ENS-Lyon, the French CNRS, Lyon 1 University, the European Research Council under the European Union’s Horizon 2020 research ADVERTISERS and innovation program (ERC 714519/HP4all and Marie Skłodowska-Curie 766402/ZULF). The NHMFL is supported by NSF/DMR-1644779 and the State of Florida, NIH S10 OD018519, NIH AUTHOR INDEX P41-GM122698-01, and NSF CHE-1229170. PO090 POSTERS / HYPERPOLARIZATION 243 ZHUORAN WANG1, DYNAMIC NUCLEAR POLARIZATION ENABLES THROUGH-BOND DAVID GAJAN,1 195PT-13C SOLID-STATE NMR CORRELATION SPECTROSCOPY OF GEORGES MENZILDJIAN,1 LUKAS ROCHLITZ,2 SURFACE-SUPPORTED PT COMPLEXES AT NATURAL ABUNDANCE DOMENICO GIOFFRÉ,2 NICOLAS KAEFFER,2 Supported single-site Pt species have recently attracted much in- species. Our approach exploits 1 J(195Pt-13C) couplings in Pt complex AMRIT VENKATESH,3,4 terest as highly efficient heterogeneous catalysts by themselves or having Pt-C bonds, as these scalar interactions are commonly large CHRISTOPHE COPÉRET,2 as precursors to generate metallic nanoparticles. In that context, while being extremely sensitive to the local structural environment. AARON ROSSINI,3,4 well-defined Pt complexes have been grafted on silica, whose co- We demonstrate this approach on two silica-anchored Pt complex- ANNE LESAGE1 ordination environment needs to be understood at the molecular es: in the first sample (Pt-NHC@SiO ) the coordination carbon has 2 1 level. Platinum-195 NMR would in principle be a method of choice been isotopically enriched while the second sample (PtMe(COD)@ High-Field NMR Center of Lyon, CNRS, ENS Lyon, UCB Lyon, Villeurbanne, France to characterize the local chemical environment and electronic SiO ) was synthesized by grafting natural abundance precursors. In 2 2Department of Chemistry and Applied structure of the metal center, providing invaluable structural in- the two cases, two-dimensional (2D) 13C{195Pt} J-HMQC spectra were Biosciences, ETH Zürich, Switzerland 3 formation to rationalize the design of the catalytic materials. 195Pt successfully acquired in short experimental times. For Iowa State University, Department of Pt-NHC@ Chemistry, Ames, IA, USA is however a low-sensitivity NMR probe as it usually experiences an SiO , the 2D 13C{195Pt} J-HMQC spectrum unexpectedly reveals two 2 4U.S. DOE Ames Laboratory, Ames, IA, USA extremely broad chemical shift anisotropy (CSA) that greatly reduc- kinds of Pt surface sites. Both the corresponding isotopic 195Pt and es the experiment sensitivity. For surface complexes, this limitation 13C chemical shifts as well as the 1 J(195Pt-13C) values for each site were adds to the low concentration of Pt sites, making any investigation determined. These two distinct sites could not be detected by pre-by 195Pt NMR extremely challenging. vious NMR investigations [2] nor by static 195Pt DNP SENS measure- ments. DFT calculations are currently under way to propose struc- INTRODUCTION We have recently demonstrated that the 195Pt CSA parameters and tural models for the two sites. A correlation between the Pt center SPONSORS the Pt−H distances of Pt surfaces sites could be determined by com- and methyl carbon is observed in the 2D 13C{195Pt} J-HMQC spectrum PROGRAM bining room temperature proton-detected NMR spectroscopy with of PtMe(COD)@SiO , providing direct evidence that the methyl PROGRAM — BY DAY dynamic nuclear polarization surface enhanced NMR spectroscopy 2 ligand persists on the surface and is bounded to Pt. The 1 J(195Pt-13C) TOPICS (DNP-SENS) [1]. Measuring correlations between the Pt center and value measured from this spectrum is in agreement with that ob- PRIZE LECTURES other neighboring NMR-active nuclei would be highly desirable to served for the molecular precursor [3]. The methodology described PLENARY LECTURES increase structural content. here represents a new step forward in the atomistic description of TUTORIAL LECTURE Here, we demonstrate that the sensitivity boost provided by DNP al- catalytically relevant surface metal complexes. INVITED AND PROMOTED LECTURES lows one to record 195Pt-13C correlations on diluted platinum surface POSTERS ADVERTISERS AUTHOR INDEX References: [1] A. Venkatesh et al., J. Am. Chem. Soc. 2020, 142 (44), 18936-18945. [2] P. Berruyer et al., J. Am. Chem. Soc. 2017, 139, 849-855. [3] P. Laurent et al., Dalton Trans. 2013, 42, 238-248. PO091 POSTERS / FIELD-CYCLING NMR RELAXOMETRY 244 MARIE BERNARDI, STUDY OF HEAVY METAL LOADED RESINS BY VUONG QUOC LAM, BENCHTOP NMR GOSSUIN YVES University of Mons, Department Heavy metals discharged by industrial wastewater to the environment has become a major public health and envi- for Biomedical Physics, Mons, ronmental concern [1]. For example, heavy metals ions such as Ni (II) and Cu (II) are known to be toxic and must be Belgium removed from wastewater. However, Ni (II) and Cu (II) ions also have paramagnetic properties which previously allowed the use of Magnetic Resonance Imaging (MRI) and Nuclear Magnetic Resonance (NMR) relaxometry to follow their migration and their adsorption on different media [2-4]. The purpose of this study is to monitor the removal of Ni (II) and Cu (II) from water by amberlite IR120 with T1 and T relaxometry. In order to obtain the adsorption 2 isotherms, different samples containing the same amount of Amberlite IR120 resin was put in contact with aque- ous solutions containing Ni (II) or Cu (II) ions at different concentrations, before being shaken by a vortex mixer. Once the equilibrium reached, the longitudinal and transversal relaxation time ( T , T ) of the solution was measured 1 2 which allowed the determination of the amount of adsorbed metal. Study of the loaded resin was afterwards carried out using a larger amount of resin which was dried and rehydrated before being analyzed. The equilibrium adsorption behavior of Ni (II) or Cu (II) can be satisfactorily described by the Langmuir model, with maximum adsorption capacity of 81.5 mg g−1 and 78.3 mg g−1 for Cu (II) and Ni (II) respectively whereas the sorption equilibrium constants are 0.98 L mg-1 (Cu (II)) and 1.8 L mg-1 (Ni (II)). The longitudinal and transverse relaxation of the wet resin are shown to be biexponential. The relaxation rate of the fast relaxing water fraction of the wet resin can be correlated with metal contents obtained by Atomic Emission Spectroscopy (ICP-AES). The next step will be to reproduce these ex- INTRODUCTION periments for other adsorbents and paramagnetic ions at different magnetic fields. With this methodology, the SPONSORS adsorption could be followed with low-cost portable NMR device. In the future, it will also be interesting to carry out PROGRAM a so-called NMR column experiment in order to investigate the adsorption within the resin in real-time. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] Tchounwou, P. B., Yedjou, C. G., Patlolla, A. K., & Sutton, D. J., Molecular, Clinical and Environmental Toxicology. Experientia Supplementum, 2021, ADVERTISERS 101, 133-164. [2] Gossuin, Y., Hantson, A.-L., & Vuong, Q. L, Journal of Water Process Engineering, 2020, 33, 101024. [3] Gossuin, Y., & Vuong, Q. L., Separation AUTHOR INDEX and Purification Technology, 2018, 202, 138-143. [4] Gusiatin, Z. M., & Kulikowska, D., Waste Management, 2014, 34, 1227–1236. PO092 POSTERS / FIELD-CYCLING NMR RELAXOMETRY 245 MORENO PASIN1, VILLIAM BORTOLOTTI2, A NOVEL FAST FIELD CYCLING APPROACH TO OBTAIN PELLEGRINO CONTE5, ANASTASIIA FIELD DEPENDENT T -T 2D MAPS NAGMUTDINOVA2, LEONARDO BRIZI3, 1 2 GERMANA LANDI4, PAOLO LO MEO6, Fast field cycling (FFC) NMR relaxometry [1] is a very valuable tool for studying molecular dynamics of many differ-DELIA FRANCESCA CHILLURA MARTINO6, ent chemical systems such as soils, food, environment [2], organic systems [3] and a number of different porous ma-VALERIA MANILLA1, GIANNI FERRANTE1 terials [3,4]. Up to now FFC NMR relaxometry has been used to describe the dependence of longitudinal relaxation 1Stelar srl, Mede, Italy time T from the relaxation field over a wide range of values spanning from ultra-low magnetic field up to a few Te-1 2University of Bologna, DICAM, Bologna, Italy 3 sla. This unique capability offered by FFC NMR relaxometry is exploited to obtain both nuclear magnetic resonance University of Bologna, DIFA, Bologna, Italy 4University of Bologna, Mathematical Department, Bologna, Italy relaxation dispersion (NMRD) profiles reporting 1/T -vs-n , and distributions of T components at different fields. 1 L 1 5University of Palermo, Department of Agriculture, Food, and Forestry Science (SAAF), Palermo, Italy So far, no information has never been obtained on the behavior of the transversal relaxation time T as affected by 2 6University of Palermo, Department of Biological, Chemical and the modulation of T when the proton Larmor frequency of the applied magnetic field is changed. In the past, the 1 Pharmaceutical Sciences and Technologies, Palermo, Italy main reason for such a lack of information was due to some hardware limitations. However, some recent develop- ments of the electronics, allow us to produce FFC NMR instruments which can be used also to measure T relaxation 2 values. In the present study, an attempt to monitor the behaviour of T as affected by T modulation at different magnetic 2 1 fields was given by computing the experimental data with a new roboust 2D NMR inversion algorithm that uses a locally adapted multi-penalty regolarization approach [5]. A cheese sample was analysed in order to obtain FFC 2D maps [6] from which more information on the different motion components can be retrieved. According to a INTRODUCTION previous study [7], at least four different components could be identified (i.e. bulk water, bound water interacting SPONSORS with the polar groups in cheese, fats, proteins). PROGRAM This study highlights a novel method for obtaining 2D maps at different magnetic fields and shows the usefulness PROGRAM — BY DAY of these maps for discriminating between different motion components in only a few shots. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] R.Kimmich & E.Anoardo, Prog. Nucl. Magn. Reson. Spectrosc. 2004, 44: 257. [2] P Conte, G. Alonzo, eMagRes 2013, 2: 389–398. [3] R.Steele & ADVERTISERS al., Magn. Res. Chem. 2016, 54: 502-509. [4] D.Faux & al. Molecules 2019, 24: 3688. [5] V. Bortolotti & al., Computational Geosciences 2021, 25:1215-1228.[6] AUTHOR INDEX N.Marigheto & al., Journal of Magnetic Resonance 187.2 (2007): 327-342. [7] P.Conte & al., Food Research International 2021, 139: 109845. PO093 POSTERS / FIELD-CYCLING NMR RELAXOMETRY 246 ANTON GRADIŠEK1, MOLECULAR DYNAMICS IN CLOSO-BORATE SYSTEMS MATHIAS JØRGENSEN2, STUDIED BY MEANS OF NMR MARK PASKEVICIUS2,3, BJARNE R. S. HANSEN2, Metal closo-borates are an emerging class of solid electrolytes, showing high thermal, chemical, and electrochem-MITJA KRNEL1, ical stability, as well as superionic conductivity in their disordered state. Generally, the compounds exist with an JANEZ DOLINŠEK1, ordered crystal structure at room temperature (RT) and undergo a thermally induced order−disorder transition, in TORBEN R. JENSEN2 which both the anion and cation sublattices become disordered as the large anion cages are distributed over two 1Jožef Stefan Institute, Ljubljana, crystallographic positions. This liquid-like environment allows the cations to easily move in the structure, resulting Slovenia 2 in a sharp, order of magnitude increase in the ionic conductivity. The conductivity is assisted by the rotations of the Interdisciplinary Nanoscience Center (iNANO), Department of Chemistry, boron cages. Aarhus University, Aarhus, Denmark Here, we present a study of molecular dynamics in three closo-borate systems, (NH ) B H , (NH ) B H , and Ag-4 2 12 12 4 2 10 10 3Department of Physics and B H , the first two being weak ionic conductors because of the large mobile ion NH +, while the third one is an 2 12 12 4 Astronomy, Fuels and Energy Technology Institute, Curtin excellent conductor [1,2]. We determined the activation energies for the reorientations of boron cages by means of University, Perth, Australia temperature-dependent NMR spin-lattice relaxation measurements on 1H and 11B and complemented them by ionic conductivity measurements. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] A. Gradišek et al., J. Phys. Chem. C. 2018, 122, 17073-17079. [2] A. Gradišek et al ., J. Phys. Chem. C. 2021, 125, 10, 5534–5541 (2021). PO094 POSTERS / FIELD-CYCLING NMR RELAXOMETRY 247 ANANT KUMAR1, PROBING MOLECULAR DYNAMICS IN THE N AND NTB PHASES OF CARLOS CRUZ1, THE TWIST-BEND NEMATOGEN DTC5C7 THROUGH MAGNETIC JOÃO L. FIGUEIRINHAS1, PEDRO J. SEBASTIÃO1, RESONANCE RELAXOMETRY CHRIS WELCH 2, GEORG H. MEHL2 The liquid crystal (LC) materials forming the Ntb phase are interest- To this aim, dispersions of relaxation rates were measured over a 1 ing not just owing to their unique phase structures and properties very broad frequency range and extensively analyzed in the nemat-IST, Universidade de Lisboa, Av. Rovisco Pais, Lisbon, Portugal but also their exploitation in many new applications [1]. Despite a ic, Ntb, and smectic phases. With this investigation, we expand the 2Dept. of Chemistry, University of decade of discovery [2], the Ntb phase remains extensively debat- current knowledge on these systems as the proton NMR relaxation Hull, Cottingham Road, Hull, UK ed over its structure and properties [3,4]. Alternative to the usual techniques give a detailed insight into physical properties such as twist-bend model [2,3], the polar-twisted (Npt) geometry has also correlation times, viscoelastic parameters and correlation lengths been endorsed for the structure of the Ntb phase [4]. Thus, neither that are signature characteristics of the structural properties of of these geometries has been fully established experimentally. Also, the studied mesophases. Our results indicate that the molecular despite a large body of studies devoted to the Ntb materials, many organization in the Ntb phase has a substantial impact on the dy-of their physical properties and molecular features influencing the namic behavior of the studied system. Significant differences were onset of the Ntb phase have not been fully conclusive. observed between the structures of the N and Ntb phases through distinct signatures of temperature- and frequency-dependent col- In this work, by means of proton nuclear magnetic resonance (1H lective mode, particularly in the low-frequency domain. Similar NMR) relaxometry, we present a molecular dynamics study of the dispersion characteristics of the Ntb and smectic phases suggest liquid crystal dimer DTC5C7. This material remains one of the rare smectic-like clustering in the Ntb phase. INTRODUCTION examples of LC materials that in addition to exhibiting nematic and SPONSORS twist-bend nematic phases also possesses a lamellar smectic phase. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1]. Y. Arakawa, K. Komatsu, J. Feng, et al. Materials Advances, 2020. [2]. V. P. Panov, M. Nagaraj, J. K. Vij, et al. Phys. Rev. Lett. 2010, 105, 167801; M. Cestari, S. Diez-Berart, D. A. POSTERS Dunmur, et al. Phys. Rev. E, 2011, 84, 031704. [3]. I. Dozov and G. R. Luckhurst. Liquid Crystals, 2020, 1-18. [4] E. T. Samulski, A. G. Vanakaras, and D. J. Photinos. Liquid Crystals, 2020, 1-6. ADVERTISERS Acknowledgements: Portuguese Science and Technology Foundation (FCT) through CeFEMA strategic project UID/CTM/04540/2013; European Union and FCT through Grant M-ERA- AUTHOR INDEX NET2/0006/2016 (CellColor) and the EPSRC (project EP/M015726/1). PO095 POSTERS / FIELD-CYCLING NMR RELAXOMETRY 248 MATTHIAS LEHMANN1, FAST FIELD-CYCLING NMR RELAXOMETRY AND STEFAN MAISCH1, LIQUID CRYSTAL BIAXIALITY NIKOLAI SCHEURING1, JOSE CARVALHO2,3, In the search of the predicted biaxial nematic phase, a series of shape-persistent board-shaped mesogens with max-CARLOS CRUZ2,3, imum molecular biaxiality and a dipole along the minor molecular axis were designed to form nematic (N) meso- PEDRO J. SEBASTIÃO2,3, phases. One compound exhibits a wide nematic temperature range, which can be supercooled to room tempera- RONALD Y. DONG4 ture. A comprehensive variable temperature X-ray study on aligned samples reveals patterns being dominated by 1Institute of Organic Chemistry, University of the form factor of very small aggregates, from which the aspect ratio of the lead compound with length (L):breadth Würzburg, Germany; (B):width (W) of 10.73:3.16:1.23 could be obtained. Variable temperature proton relaxation studies on this mesogen matthias.lehmann@uni-wuerzburg.de 2Centro de Física e Engenharia de Materiais were carried out over a wide frequency range. Avançados, Lisbon, Portugal; pedro.jose.sebastiao@tecnico.ulisboa.pt The global fit of the frequency dispersions at five temperatures with a motional model requires in addition to the 3Department of Physics, Instituto Superior usual rotation/reorientation contribution, two independent director fluctuations contributions: one for the con-Técnico, Universidade de Lisboa, Lisbon, Portugal ventional nematic order director (n) fluctuations and the other for the minor director (m) fluctuations (normal 4Department of Physics and Astronomy, to n). The correlation length of the minor directors determined by NMR could extend to 5–8 molecules in the W University of British Columbia, Vancouver, direction, but only to the nearest neighbour in the B direction, as found by X-ray diffraction. Both X-ray and NMR BC, Canada; rondong@phas.ubc.ca studies indicate that these new types of lead structure are extremely promising to find the long sought-after biaxial N mesophase [1]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] M. Lehmann, S. Maisch, N. Scheuring, J. Carvalho, C. Cruz, P. J. Sebastião, R. Y. Dong, Soft Matter 2019, 14, 84896. POSTERS Acknowledgements: COST Action CA15209 European Network on NMR Relaxometry. Portuguese Science and Technology Foundation (FCT) through CeFEMA ADVERTISERS strategic projects UID/CTM/04540/2013 and UID/CTM/04540/ 2019; European Union through Grant FP7-PEOPLE-2007-1-1- ITN-215884 (DENDREAMERS); AUTHOR INDEX and European Union and the Portuguese Science and Technology Foundation (FCT) through Grant M-ERA-NET2/0006/2016 (CellColor). PO096 POSTERS / MRI IN MATERIAL SCIENCE AND BIOMEDICAL APPLICATIONS 249 ANNA MORALES- ZN INCORPORATION IN SYNTHETIC C-S-H AND ITS MELGARES1,2*, EFFECT ON CEMENT HYDRATION THROUGH DNP PINELOPI MOUTZOURI1, AMRIT VENKATESH1, ENHANCED MAS NMR PAUL BOWEN2, KAREN SCRIVENER2, The contribution of CO emissions from cement production account for 8-10% of the total emissions we produce. 2 LYNDON EMSLEY1 One possible solution is the partial substitution of the clinker by Supplementary Cementitious Materials (SCMs). 1 These are materials which production does not imply the formation of CO , but tend to lower the early-age strength École Polytechnique Fédérale de 2 Lausanne, Laboratory of Magnetic of Portland cement. However, it has recently been demonstrated that the addition of Zn can enhance the mechan- Resonance, Lausanne, Switzerland ical strength of a clinker. 2École Polytechnique Fédérale de Lausanne, Laboratory of The goal of this project is to understand the role of Zn in synthetic single-phase C-S-H [1] by means of solid state Construction Materials, Lausanne, nuclear magnetic resonance (NMR). Results show evidence of Zn incorporation into the C-S-H structure and in- Switzerland sight about the exact position of Zn at the atomic level. DNP enhanced Solid state 29Si NMR CP experiments show two clear trends. Firstly, the intensity of the peak which corresponds to the Q2 sites increases as the Zn content is increased; and secondly, a new peak around -70ppm becomes more evident as more Zn is added to the sample too. These two facts verify the hypothesis that Zn is inside the C-S-H structure since it is changing the chemical environment of some Si sites and changes the ratio between Q1 sites (which correspond to Si at the end of a silicate chain) and Q2p (which correspond to Si which are adjacent to a Q1 Si and a bridging Si). Incredible Natural Abundance Double-Quantum Frequency Transfer Experiments (INADEQUATE) have also been INTRODUCTION done in order to determine how the relation between Q species populations evolve as more Zn is incorporated into SPONSORS the system. Results show that the addition of this element promotes longer chains at the atomic scale. PROGRAM PROGRAM — BY DAY Understanding the mechanisms by which Zn is incorporated into the C-S-H structure will be vital in order to be able TOPICS to add this element into the industrial process with the ultimate goal of reducing the CO2 emissions of the cement PRIZE LECTURES industry. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Scrivener, K. et al. Cement and Concrete Research 2019, 124. AUTHOR INDEX Acknowledgements: Pinelopi Moutzouri, Amrit Venkatesh, Manuel Cordova, Ziga Casar, Aslam Kuhni, Davide Demurtas, Maya Harris and Andrea Teixeira. PO097 POSTERS / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 250 ADITYA MISHRA, SUPRAMOLECULAR STRUCTURE OF LAYERED MICHAEL A. HOPE, HYBRID PEROVSKITES DETERMINED USING NMR PARAMVIR AHALAWAT, TORU NAKAMURA, CRYSTALLOGRAPHY MANUEL CORDOVA, DOMINIK J. KUBICKI, Here, we show how multinuclear (1H, 19F and 13C) solid-state NMR in combination with molecular dynamics and FARZANEH JAHANBAKHSHI, chemical shift calculations can be employed to determine the complete atomic-scale molecular structure of the MARKO MLADENOVIĆ, organic spacer cations in layered hybrid perovskites [1]. Solid-state NMR has recently emerged as a powerful tech-URSULA ROETHLISBERGER, nique to study the structure and dynamics of hybrid organic–inorganic perovskites. NMR is exquisitely sensitive to JOVANA V. MILIĆ, chemical information such as cation incorporation, halide mixing, phase segregation and dynamics [2,3]. Layered MICHAEL GRÄTZEL, perovskites, which comprise slabs of the perovskite structure separated by a hydrophobic organic spacer layer [4], LYNDON EMSLEY have shown potential to improve the stability of perovskite solar cells, but they suffer from poor crystallinity and conductivity which impair photovoltaic efficiency. Supramolecular engineering [2,5] is a promising approach to Institut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de improve the electronic property and crystallinity by harnessing non-covalent interactions to direct the structure of Lausanne (EPFL), Lausanne, Switzerland the organic spacer layer. However, to assess this structure directing effect the complete atomic-level structure must be determined, which was not previously possible. NMR is ideally suited to study hybrid materials. Here, chemical shift-based NMR crystallography was used to determine the structure of the spacer layer in Ruddlesden–Popper perovskites featuring a bilayer of monovalent organ-ic spacer cations, in this case phenylethylammonium (PEA+), pentafluorophenylethylammonium (FEA+), or a 1:1 INTRODUCTION mixture of the two. Double resonance NMR experiments demonstrate the proximity of the two aromatic systems; SPONSORS however, comparison of chemical shifts obtained from candidate structures with experiment indicates that a phase PROGRAM segregated structure most closely matches the experimental NMR parameters. Taken together, this shows the pres- PROGRAM — BY DAY ence of a nano-scale phase segregated structure. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] M.A. Hope et al., J. Am. Chem. Soc. 2021, 143, 356. [2] J.V. Milić et al., Adv. Energy Mater. , 2019, 9, 1900284. [3] D.J. Kubicki et al., J. Am. Chem. Soc. AUTHOR INDEX 2017, 139, 10055. [4] I.C. Smith et al., Angew. Chem. , 2014, 126, 11414. [5] M.A. Ruiz-Preciado et al., J. Am. Chem. Soc. 2020, 142, 1645. PO098 POSTERS / ENERGY STORAGE AND CONVERSION MATERIALS, CATALYSTS 251 JANEZ VOLAVŠEK1,2, LOCAL STRUCTURAL TRENDS AFTER DIFFERENT TEMPERATURE NATAŠA ZABUKOVEC CALCINATION OF AL O AND CU-AL O CATALYST SUPPORTS: LOGAR1,3, 2 3 2 3 TADEJ ŽUMBAR1,4, INSIGHT BY SOLID STATE NMR NATAŠA NOVAK TUŠAR1,3, GREGOR MALI1,2,3 In the field of catalysis aluminium oxide, Al O , is often used as By studying 27Al ssNMR spectra we observed that by increasing cal-2 3 1 catalyst support, which can be modified by distinct preparation cination temperature the local structure around Al atoms gets more National Institute of Chemistry, Department of Inorganic Chemistry and methods such as different synthesis routes, additives and calcina- ordered – peaks get narrower and take more distinct quadrupolar Technology, Ljubljana, Slovenia tion temperatures. Such variation in preparation may lead to con- shape. A parallel series of supports with Cu as dopant have the same 2University of Ljubljana, Faculty of trolled manipulation of coordination number and structural order general temperature trend. We also find that supports with Cu ad-Mathematics and Physics, Ljubljana, Slovenia around aluminium atoms, and consequently enhanced catalytic ditive, calcined at temperatures up to 1000 °C, have slightly more 3University of Nova Gorica, Nova Gorica, properties for specific degradation processes of volatile organic tetrahedral coordinated Al compared to those without added Cu. Slovenia 4University of Ljubljana, Faculty of compounds [1]. Interestingly, calcination at 1200 °C is an exception, where addition Chemistry and Chemical Technology, of Cu significantly changes the local structure of Al atoms. Addition Ljubljana, Slovenia Synthesis of catalyst supports presented here was based on alumi- of Cu at highest calcination temperature of 1400 °C has no influence na pseudoboehmite powder, which was calcined in oxidative at- on Al environment, where we are left with only one type of tetra- mosphere, ensuring highest oxidation state of the material. In this hedral coordinated Al sites. To investigate if the operation of the preliminary study we investigated the local structure of aluminium catalyst support influences the Al local structure, we performed a atoms by using solid state nuclear magnetic resonance spectrosco-degradation test of toluene at 200 °C, and found out that the Al O py (ssNMR). From a series of Al O catalyst supports calcined at dif- 2 3 2 3 support is stable, meaning we did not observe any difference in 27Al INTRODUCTION ferent temperatures, ranging from 800 to 1400 °C, we reveal some ssNMR spectra before and after use of said catalyst support. SPONSORS of the temperature dependent trends regarding the local surround- PROGRAM ing of Al atoms. In parallel we also looked at the influence of copper PROGRAM — BY DAY additive on Al coordination at various calcination temperatures. In TOPICS all these samples 8 weight % of Cu relative to the Al O support was 2 3 PRIZE LECTURES used. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] Y. Xie, D. Kocaefe, Y. Kocaefe, J. Cheng, W. Liu, Nanoscale Res Lett. 2016, 11, 259-269. AUTHOR INDEX Acknowledgements: Slovenian Research Agency (ARRS). PO099 POSTERS / ORGANIC AND COMPOSITE SOLIDS 252 AXEL GANSMÜLLER1, INSIGHT INTO TRANS-[Ru(NO)(py) F](ClO )2 PHOTO- 4 4 ARTEM MIKHAILOV2, COMMUTATION PROPERTIES BY SOLID STATE NMR GENNADIY KOSTIN2, JESUS RAYA3, AND DFT CALCULATIONS CYRIL PALIN1, THEO WOIKE1, Photoinduced isomers can be generated in a variety of compounds, each having its specific structural response DOMINIK SCHANIEL1 and wavelength sensitivity. This gives rise to interesting properties such as photochromism and photorefractivity 1 for holographic data storage and optical switches. Fundamental questions in this context are to what extent the CRM2, UMR 7036 CNRS, Université de Lorraine, Nancy, France electron density distribution in the photo-generated isomer has changed with respect to the ground state config-2Nikolaev Institute of Inorganic uration, as well as how the structural and electronic characteristics of the photoswitches are linked to the thermal Chemistry, Siberian Branch of the stability and optical properties of the photoproducts. Russian Academy of Sciences, Novosibirsk, Russian Federation 3 In this communication we present solid-state NMR results obtained on the complex trans-[Ru(py) (15NO)F](ClO ) , Institut de Chimie, UMR 7177 4 4 2 CNRS, Université de Strasbourg, where the quasi-fourfold axis F-Ru-15N-O is nearly linear in the ground state and the unit cell contains two crys-Strasbourg, France tallographic independent molecular units. Upon light irradiation (420nm), two metastable linkage isomers can be generated, one by rotating NO by about 90° (MS2) and one by an inversion of the NO ligand by 180° (MS1) [1]. We show here the successful low temperature trapping and SSNMR observation of the diamagnetic light induced metastable state MS1. In order to gain insight into the bonding properties and changes of the electron density upon photo-isomerization, we compare the 15N and 19F solid-state NMR results with those obtained from DFT calculations based on our recent photo-crystallographic X-ray diffraction study [2]. These results are then interpreted based on INTRODUCTION local changes of the electron density of the atoms along the F-Ru-NO axis. SPONSORS Interestingly, we observe different commutation properties within the same crystal for the two inequivalent cations PROGRAM and supply an explanation based the interaction of the cation with it’s counterion. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] G. A. Kostin, A. A. Mikhailov, N. V. Kuratieva, D. P. Pishchur and A. N. Makhinya, New Journal of Chemistry, 2018, 42, 18928–18934, [2] A. A. AUTHOR INDEX Mikhailov, E. Wenger, G. A. Kostin and D. Schaniel, Chemistry – A European Journal, 2019, 25, 7569–7574. PO100 POSTERS / ORGANIC AND COMPOSITE SOLIDS 253 SAVVAS ORFANIDIS1,3, SOLID-STATE NMR AS AN INNOVATIVE NON-DESTRUCTIVE M. RAIMONDO2, EVALUATION METHODOLOGY IN COMPOSITE SELF-HEALING L. GUADAGNO2, A.S. PAIPETIS3, SYSTEMS FOR AEROSPACE ENGINEERING APPLICATIONS. M. FARDIS1, G. PAPAVASILLIOU1 The self-healing polymer composites have gained increasing re- The self-healing system was tested in different conditions in order 1 search interest over the past several decades and the technological to understand the behavior of the self-healing process, we examine Institute of Nanoscience & Nanotechnology, NCSR interest of the most demanding industries such as aerospace, auto- the polymerization progress at three different temperatures (25oC, Demokritos, Attiki, Greece motive and ship building industries [1]. The researches aim to find 40 oC, 60 oC) those temperatures were chosen due to operation en-2Department of Industrial an efficient healing mechanism that can lead to an extended life vironment of aerospace applications and we examine two differ-Engineering, University of Salerno, Fisciano (SA), Italy; cycle of the structure [2][3]. This study focused on the development ent epoxy-diluent mixtures (10%,25%) in order to observe how lguadagno@unisa.it (L.G.) of an innovative non-destructing evaluation methodology by taking the presence of diluent effect the polymerization progress. 2D 1H 3Department of Materials Science & Engineering, University of Ioannina, the advantage of a powerful spectroscopic technic such as Nuclear NMR diffusion–relaxation D–T2 measurements were performed to Greece Magnetic Resonance (NMR) and transform it into a non-destructing monitor the diffusion properties of the polymerization reactions of evaluation tool. In this study we examine the rheological behavior the self-healing agent during the self-healing progress of the dam-and the polymerization mechanisms of the self-healing agent af- aged structure. The final step of this study was to simulate the final ter the damage occurs. The examined structure of this scenario is self-healing system, the coating with the dispersion of micro-capa protective coating (Aerowave 3003 epoxy primer) with dispersed sules and hardener then we damaged the system and we monitored self-healing microcapsules and catalyst Curing Solution 6007. The the diffusion properties of the system during a thermal scanning enclosure of the self-healing microcapsules is a mixture of an ep- (25oC, 40 oC, 60 oC) and we achieved to monitor the consumption of INTRODUCTION oxy part, a diluent and a chromophore (DGEBA+ BDE+ Solvent Red the hardener after he damage occurs. All the above experimental SPONSORS 242). Our approach is to mapping the polymerization reactions procedures were performed in the stray field of a 4,7 T Bruker super- PROGRAM during the self-healing process in real time by monitoring the T2 conductive magnet providing a 34,7 T/m constant magnetic field PROGRAM — BY DAY relaxation time and visualizing the results in polymerization maps. gradient at 1H NMR frequency of 101,324 MHz. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] D.G. Bekas, D. Baltzis, A.S. Paipetis, Materials and Design 2017, 116, 538–544. [2] Maria Kosarli, Dimitrios G. Bekas, Kyriaki Tsirka, Dimitrios Baltzis, Dimitrios T. Vaimakis- ADVERTISERS Tsogkas, Savvas Orfanidis, Georgios Papavassiliou, Alkiviadis S. Paipetis, Composites Part B 2019, 171, 78–86. [3] Ida Palazzo, Marialuigia Raimondo, Giovanna Della Porta, Liberata Guadagno, AUTHOR INDEX Ernesto Reverchon, Journal of Industrial and Engineering Chemistry 2020, 90, 287-299. PO101 POSTERS / ORGANIC AND COMPOSITE SOLIDS 254 BENJAMIN KOHN, CONFORMATION AND PACKING OF POLYANIONS IN CARLOIN NAAS, POLELECTROLYTE COMPLEX COACERVATES – UWE LAPPAN, ULRICH SCHELER A COMBINED PFG AND SOLID-STATE NMR STUDY Leibniz-Institut für Polymerfoschung Dresden e.V., Polyelectrolyte complex coacervates are formed mixing charged polymer, polycation and polyanion in solution. Dresden, Germany Such complexes find wide application in water treatment for flocculation or for controlled drug release. The conformation of polyelectrolytes in solution depends on the repelling force from the charges along the polymer chain. For a weak polyanion poly(maleic anhydrite-co-ethylene) the depends on pH and ionic strength. PFG NMR offers measures for both conformation and charge of polyelectrolytes in solution. The effective charge is inferred from electrophoresis NMR while diffusion NMR yields the hydrodynamic size as a measure for the conformation. With increasing pH the weak polyacid dissociates generating more charges and thus a more stretched conformation. The degree of dissociation of the solid materials is determined from the relative acid proton signal intensity in 1H MAS spectra. Separating 1H spectra in two-dimensional single-quantum-double-quantum correlation spectra distinguishes between acid protons hydrogen bonded to other acid protons from others and thus identifies polyanion-rich regions in solid polymers or complexes. Two-dimensional integration of the on-diagonal and off-diagonal signals of the acid protons enables quantification. Apparently the conformation of the polyelectrolytes in the parent solutions from which the complexes are formed is partially retained in the complexes. At low pH (weak charge) this are reduced by a factor of three in the complexes at higher pH (high nominal charge) with a more stretched con- INTRODUCTION formation almost none acid-acid contacts are found in the complexes. As the pH is adjusted by NaOH, 22Na proides SPONSORS additional insight. In the complexes exhibiting ion pairs between polyanion and polycation signals from NaCl are PROGRAM found while the rest shows signals from maleate. PROGRAM — BY DAY Partially dissociated poly(maleic anhydrite-co-ethylene) exhibits strong down-field shifts for the reamining acid TOPICS protons up to 20 ppm. This is confirmed for maleic acid at various pH values. The strong hydrogen bonds are seen PRIZE LECTURES in DFT calculations using CASTEP. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX PO102 POSTERS / METHODS DEVELOPMENT 255 MARC AZAGRA1, AMMONIUM QUANTIFICATION (AQUA) FRANCESCO DE CHIARA1, IN HUMAN PLASMA BY NMR FOR CLINICAL STAGING MARTINA PEREZ2, EMMA AVITABILE2, OF FATTY LIVER JOAN-MARC SERVITJA3, LAURA BRUGNARA3, Fatty liver disease (FLD) is a common disease with identifiable markers. However, there is a lack of tools powerful enough ELISA POSE2, to provide a reliable diagnosis for FLD without requiring a biopsy [1]. It has been demonstrated that there is a direct rela-JAVIER RAMON-AZCÓN1,4, tionship between FLD and hyperammonemia in plasma because of the downregulated activity of the urea cycle enzymes IRENE MARCO-RIUS1* [2]. The physiological ammonia level in blood is constantly maintained around 40 µM and, at physiological pH (pH ≈ 7.4), 1 around 98% of ammonia is in ionic form (NH +). Pathological conditions such as liver dys-function may disrupt the me-Institute for Bioengineering of Catalonia, 4 Barcelona Institute of Science and tabolism of NH and lead to a life-threatening increase in blood [3]. 3 Technology, Barcelona, Spain 3Liver Unit, Hospital Clinic, Faculty of Here we show how proton nuclear magnetic resonance (1H-NMR) spectra of human plasma can correctly diagnose Medicine and Health Sciences, University the presence of FLD with the proper ammonia quantification. The procedure has been optimized testing different of Barcelona, Barcelona, Catalonia, Spain 4 deuterated solvents and pHs in order to achieve an accurate and realible quantification. The study has been per-Institut d’investigacions Biomèdiques Agust Pi i Sunyer (IDIBAPS), Barcelona, formed with alcoholic fatty liver disease (AFLD) and non-alcoholic steatohepatitis patients (NASH). Our studies Catalonia, Spain show for the first time a technique capable of providing an accurate and rapid diagnosis of FLD without the need 5ICREA-Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain for an invasive biopsy. This technique can be used in a clinical setting either in population screening or to allow effective targeting of treatment. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] D. R. Aldridge, E. J. Tranah, and D. L. Shawcross, J. Clin. Exp. Hepatol. , 2015, 5, S7–S20. [2] F. De Chiara, S. Heebøll, G. Marrone, C. Montoliu, S. INVITED AND PROMOTED LECTURES Hamilton-Dutoit, A. Ferrandez, ... & K.L. Thomsen,. Journal of hepatology, 2018, 69(4), 905-915. [3] R. J. Barsotti, J. Pediatr. , 2001, 138. POSTERS Acknowledgements: We thank the Barcelona Nuclear Magnetic Resonance Laboratory, in particular Francisco Cárdenas. This work is part of a project that ADVERTISERS has received funding from the European Union’s Horizon 2020 Future and Emerging Technologies Open under grant agreement 863037 (BLOC) and the AUTHOR INDEX Junior Leader Postdoctoral Fellowship Programme from “la Caixa” Banking Foundation (LCF/BQ/PI18/11630020). PO103 POSTERS / METHODS DEVELOPMENT 256 RACHA BAYZOU1, INDIRECT NMR DETECTION OF NUCLEI SUBJECT TO JULIEN TRÉBOSC1, LARGE ANISOTROPIC INTERACTIONS VIA PROTONS IVAN HUNG2, ZHEHONG GAN3, USING T-HMQC SEQUENCES OLIVIER LAFON1,3, JEAN-PAUL AMOUREUX1,4,5 Solid-state NMR is a unique technique for the characterization of the atomic-level structure and dynamics of mate-1 rials. Nevertheless, this technique can often be challenging when the investigated materials contain S nuclei subject Univ. Lille, CNRS, Centrale Lille, Unité de Catalyse et Chimie du Solide, Lille, France to large anisotropic interactions, such as chemical shift anisotropy (CSA) or quadrupolar coupling for S ≥ 1. Recently, 2National High Magnetic Field Laboratory, a through-space HMQC variant using the TRAPDOR (TRAnsfer of Populations in DOuble-Resonance) recoupling, Tallahassee, FL, USA 3 called T-HMQC, has been successfully applied for the indirect observation via protons of quadrupolar isotopes such Institut Universitaire de France, Paris, France as 14N ( S = 1) or 35Cl ( S = 3/2) [1,2]. This simple sequence is made of only four rectangular pulses: one p/2 and one p 4Riken NMR Science and Development pulses forming a spin-echo on the 1H channel and two symmetrical long pulses applied to the S spin, which rein-Division, Yokohama, Kanagawa, Japan 5Bruker Biospin, 34 rue de l’industrie, troduce the 1H- S dipolar couplings and transfer coherences between 1H and S isotopes. Due to the absence of any Wissembourg, France. recoupling scheme on the 1H channel, the T-HMQC sequence is robust to spinning speed fluctuations, and hence eliminates the t -noise. Additionaly, its sensitivity is higher than the conventional D-HMQC. 1 In the present work, we have analyzed with spin dynamics simulations and experiments the efficiency and robust- ness of the T-HMQC sequence at fast MAS for the indirect detection of 195Pt, 14N and 35Cl isotopes [3]. For 195Pt isotope, the method is robust to offset, and its efficiency increases for increasing CSA values and is still significant for values of 3 MHz. For quadrupolar nuclei, we demonstrate that the maximum sensitivity is achieved for a moderate rf-field INTRODUCTION and an offset equal to the MAS frequency. In the case of 14N nuclei, the T-HMQC methods can select the 1Q or 2Q SPONSORS coherences during t period. The selection of 2Q coherences enhances the resolution. For 35Cl isotope, the selection 1 PROGRAM of 2Q or 3Q coherences during t period improves the resolution by a factor of 18 or 3.9 with the respect to D-HMQC 1 PROGRAM — BY DAY sequence, respectively. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] I. Hung, P. Gor’kov, Z. Gan, J. Chem. Phys 2019, 151, 154202. [2] I. Hung, Z. Gan, J. Phys. Chem. Lett 2020, 11, 4734-4740. [3] R. AUTHOR INDEX Bayzou, J. Trébosc, I. Hung, Z. Gan, O. Lafon, J.-P. Amoureux, in preparation. PO104 POSTERS / METHODS DEVELOPMENT 257 MATTHIAS BRETSCHNEIDER1, MULTIQUANTUM COUNTING OF TRITYL RADICALS PHILLIP E. SPINDLER1, OLGA YU. ROGOZHNIKOVA2, Multi-Quantum coherence (MQC) has been used in NMR [1, 2] to count coupled spin in clusters. In EPR DQC experi- DMITRY V. TRUKHIN2, ments, first introduced by the Freed group [3], are used to selectively detect the dipolar interaction within a coupled BURKHARD ENDEWARD1, electron spin pair. Here we show that the number of coupled electron spins can also be obtained for larger spin ANDREY A. KUZHELEV2, clusters using this method [4]. This approach has the potential to determine the oligomeric state of spin-labeled VICTOR M. TORMYSHEV2, proteins complexes in their native environment. ELENA BAGRYANSKAYA2, To count the number n of dipolar coupled spins, experiments which selectively filter the n-quantum coherences are THOMAS F. PRISNER1 proposed. For this experiment a high accuracy in the phases of the microwave pulses and a sufficient broad band-1Institute of Physical and Theoretical width to homogeneously excite the electron spins is mandatory. This is achieved using a home built X-band EPR Chemistry and Center of Biomolecular setup based on an arbitrary wave form generator. The trityl radicals have narrow spectral width so that quantitative Magnetic Resonance, Goethe University Frankfurt, Frankfurt am Main, Germany and precise excitation of all spins by the pulses is possible. Here we demonstrate on a series of multi-trityl model 2Novosibirsk Institute of Organic Chemistry, compounds that MQ-filtered EPR experiments allow to determine the number of coupled electron spins. The trans- Novosibirsk, Russia versal relaxation times of higher quantum coherences are also measured. More common spin label for proteins are nitroxides. This spin labels have much broader EPR spectral width in comparison to trityl radicals, exceeding the excitation bandwidth of rectangular microwave pulses. Our first experimental results using broadband microwave excitation pulses on multi-nitroxide model compounds will be shown. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] G. Bodenhausen, R. L. Vold, R. R. Vold, J. Magn. Reson. 1980, 37 (1), S. 93–106. [2] M. Munowitz, A. Pines, Science 1986, 233, 525–531. [3] P. P. ADVERTISERS Borbat, J. H. Freed, Chem. Phys. Lett. 1999, 313, 145–154. [4] M. Bretschneider, P. E. Spindler, O. Yu. Rogozhnikova, D. V. Trukhin, B.Endeward, A. A. Kuzhelev, AUTHOR INDEX E. Bagryanskaya, V. M. Tormyshev, T. F. Prisner, J.Phys.Chem.Lett. 2020, 11, 6286−6290. PO105 POSTERS / METHODS DEVELOPMENT 258 MATÍAS CHÁVEZ, A GENERALIZED FLOQUET TREATMENT FOR PULSE MATTHIAS ERNST SEQUENCE OPTIMIZATION Physical Chemistry, ETH Zürich, Zürich, Switzerland Floquet theory is one of the main methods to analyze time-dependent Hamiltonians. In the standard form, Floquet theory requires a periodic Hamiltonian and does not take into account the finite length of typical rf-irradiation schemes [1]. We have developed a generalized Floquet treatment for magnetic resonance that is based on a frequency-domain representation of the Hamiltonian. The new treatment does not require a periodic time-dependent Hamiltonian and takes the finite length of pulse sequences into account. In contrast to the standard Floquet treatment that is described by Fourier coefficients at integer multiples of the basic frequency, the new treatment is based on a Hamiltonian that is defined on a continuous frequency axis. Homonuclear recoupling schemes under MAS received a great deal of attention since they can provide data for distance restrains in biomolecules. Proton-drive spin diffusion (PDSD) is one of the most important sequences to obtain distance restraints [2]. MIRROR and AM-MIRROR recoupling are tunable PDSD-based sequences that allow broadband and band-selective recoupling even at high MAS frequencies [3-5]. We present a refinement of the band-selective second-order MIRROR recoupling scheme, which allows targeted distribution of the polarization transfer, by creating side-bands in the zero-quantum spectrum. This development was enabled by the generaliza- tion of the Floquet theory described above, which provides a bijective mapping between the side-band pattern and the pulse scheme. This formalism does not require periodicity of the Hamiltonian and offers a simple route to gain analytical insight into finite-length arbitrary pulse irradiation schemes. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] I. Scholz, J. D. van Beek, M. Ernst, Nucl. Magn. Reson. 2010, 37, 39–59 [2] A.Grommek, B. H. Meier, and M. Ernst, Chem. Phys. Lett. 2006, 427, ADVERTISERS 404-409 [3] I. Scholz, M. Huber, T. Manolikas, B. H. Meier, M. Ernst, Chem. Phys. Lett. 2008, 460, 278-283 [4] J. J. Wittmann, L. Hendriks, B. H. Meier, M. Ernst, AUTHOR INDEX Chem. Phys. Lett. 2014, 608, 60–67. [5] J. J. Wittmann, V. Agarwal, J. Hellwagner, A. Lends, R. Cadalbert, B. H. Meier, M. Ernst, J. Biomol. NMR 2016, 66, 233-242. PO106 POSTERS / METHODS DEVELOPMENT 259 DARIUSZ GOŁOWICZ1,2, FAST ACQUISITION OF TOCSY TRANSFER CURVES TO ALEXANDRA ASSIST THE ANALYSIS OF UNLABELED PEPTIDES SHCHUKINA2, KRZYSZTOF The unlabeled peptides at low concentrations are usually studied by employing homonuclear 1H experiments. One KAZIMIERCZUK1 of the routine experiments performed in such studies is 2D 1H TOCSY. The TOCSY experiment carries crucial infor-1University of Warsaw, Centre of mation on the residue-type based on correlations observed in the residue-specific spin-system. The cross-peaks for New Technologies, Warsaw, Poland 2 individual residues usually appear as an easily recognizable pattern with typical chemical shifts. Sometimes, how-University of Warsaw, Faculty of Chemistry, Warsaw, Poland ever, the cross-peaks pattern is difficult to recognize, e.g. due to the peak overlap. We present that TOCSY transfer curves may serve as an extra dimension to resolve potential ambiguities in the peak assignment process. The acquisition of a series of 2D TOCSY experiments to obtain the TOCSY pseudo-dimension is very lengthy. In our study, we recorded a series of twenty-one 2D 1H TOCSY experiments with different settings of spinlock duration for each of two unlabeled peptides (Repeat-3 and Repeat-4 domains of TAU). Using this data, we evaluated if applying non-uniform sampling allows us to reduce experiment time without significantly affecting the quality of TOCSY transfer curves. To improve the analysis of the TOCSY dimension, we simulated thousands of TOCSY transfer curves for protons in peptides and proteins employing Spinach [1] and data from Protein Data Bank [2] and used them for reference if necessary. Combining the experimental and simulated data, we present that measuring the TOCSY-transfer dimension im- proves the analysis of unlabeled peptides. Moreover, the data can be required in a reasonable time thanks to the INTRODUCTION NUS. The approach provides increased reliability of the peak assignment and seems to be especially suited for stud- SPONSORS ies carried at the lower magnetic fields, where resolution may be insufficient. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] H.J. Hogben, M. Krzystyniak, G.T.P. Charnock, P.J. Hore, I. Kuprov, J. Magn. Reson. 2011, 208, 179-194. [2] H.M. Berman, J. Westbrook, Z. Feng, G. ADVERTISERS Gilliland, T.N. Bhat, H. Weissig, I.N. Shindyalov, P.E. Bourne, Nucleic Acid Res. 2000, 28, 235-242. AUTHOR INDEX Acknowledgements: Authors thank National Science Centre (Poland) for support with grant PRELUDIUM 2019/33/N/ST4/02751. PO107 POSTERS / METHODS DEVELOPMENT 260 ARNAUD DI BITETTO1,2, PROBING INTERLAYER ANIONIC STRUCTURES BY GWENDAL KERVERN2, SOLID-STATE NMR IN NI/AL PARAMAGNETIC LAYERED ERWAN ANDRÉ1, CÉDRIC CARTERET1 DOUBLE HYDROXYDES 1Laboratoire LCPME — UMR 7564 (UL/CNRS), Villers-lès-Nancy, France Many recent solid-state NMR studies have presented various results on structural and dynamic aspects in the chem-2CRM2 — UMR 7036 (UL/CNRS), istry of Layered Double Hydroxide (LDH) materials. Some of the most recent investigations, based on recent inno-Faculté des Sciences et Technologies, vations, demonstrated deep insights into the structure of the cationic part of Mg/Al-based LDH materials, as well as Université de Lorraine, Vandoeuvre-lès-Nancy CEDEX, France the layer/interlayer interface[1,2,6]. The influence of synthesis conditions on the interlayer anion composition has been demonstrated as well as dynamic information inferred from relaxation studies on anionic species [3-5]. In this presentation, we propose to show how we used a different set of metal centres, including paramagnetic transition metals in the hydroxide layers in order to take advantage of the paramagnetic interactions between electrons and nuclei and probe some intriguing structural features in nitrate LDH that show X-Ray characteristics as well as dynamical behaviour much different than that of their carbonated equivalent[7]. We implemented a model for fast-prediction of NMR spectra in paramagnetic species. Since anions in the interlayer space are not bound to cations in the hydroxide layers, we reduce the calculation of the hyperfine interaction in paramagnetic species to its dipolar component. By calculating this dipolar interaction and superimposing it on the chemical shift tensor measured on diamagnetic equivalents of carbonate and nitrate LDH, we can predict the spectra and measure the relative orientation of the diamagnetic and paramagnetic chemical shift tensors. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] Cadars, S. et al. Chem. Mater. 2011, 23, 2821–2831. [2] Sideris, P. J. et al. Science 2008, 321, 117, 113. [3] Reinholdt, M. X. et al. J. Phys. Chem. C 2009, ADVERTISERS 113, 3378–3381. [4] Reinholdt, M. X. et al. J. Phys. Chem. C 2009, 113, 10623–10631. [5] Reinholdt, M. X. et al. J. Chem. Mater. 2006, 18, 2567–2576. [6] Zhao et al. AUTHOR INDEX et al. Adv. Funct. Mater. 2014, 24, 1696–1702. [7] Di Bitetto et al. J. Phys. Chem. C. 2017, 121, 7276-7281 PO108 POSTERS / METHODS DEVELOPMENT 261 KRISTINA S. LIU1, SURFACE NMR USING QUANTUM SENSORS IN DIAMOND ALEXANDER HENNING2, MARKUS W. HEINDL1,2, This work reports the experimental demonstration of NMR-spec- common support in catalysis and materials science and thus rep- ROBIN D. ALLERT1, troscopy at functional planar surfaces using a new technique called resents a scientifically and industrially relevant functional interface JOHANNES. D. BARTL2,3, “surface NV-NMR”. With this approach, we successfully realize a for demonstrating our method [4]. This surface is then functional-IAN. D. SHARP2, longstanding goal in surface and interface science, with immedi- ized by means of phosphonate chemistry to form self-assembled ROBERTO. RIZZATO1, ate relevance to catalysis, material, and bioanalytical research [1]. monolayers. Surface NV-NMR is not only able to detect these or-DOMINIK. B. BUCHER1 Characterizing the molecular properties of surfaces under ambient ganic monolayers but also to monitor the surface chemistry in real 1 and reactive conditions with non-invasive probes is a fundamental time during molecular self-assembly at the solid-liquid interface. Department of Chemistry, Chair of Physical Chemistry, Technical University scientific challenge. NMR is a non-invasive, molecular level spec- The use of a diamond-based sensor that is chemically inert and of Munich, Garching bei München, troscopic technique and is ideally suited for this purpose but lacks can withstand high temperatures and high pressures will not only Germany 2 the sensitivity to probe the small number of spins at planar surfaces bridge the “pressure gap” in surface science but has the resilience Walter Schottky Institute and Physics Department, Garching bei München, [2]. Here, we demonstrate a novel method based on diamond to probe chemical reactions even under harsh conditions in-situ [5]. Germany quantum technology [3] that is applied to overcome this The functionality of surface NV-NMR under chemically relevant 3Department of Chemistry, WACKER-Chair for Macromolecular Chemistry, long-standing fundamental limitation of NMR, thereby pro- conditions and with low technical complexity makes it an advanta- Technical University of Munich, viding a new tool for chemical analysis at surfaces. This tech- geous technique for surface science (i.e. not just under vacuum, but Garching bei München, Germany nique is based on the utility of quantum sensors in diamond and also at active solid/gas and solid/liquid interfaces). Our technique demonstrates their capability for performing NMR at surfaces and introduces NMR at surfaces and is a key step towards wide-ranging interfaces under ambient conditions. We use atomic layer deposi-applications for in-situ analysis of catalysis, materials, biological, tion to prepare thin Al O films on the diamond. This material is a and 2D materials research. 2 3 INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] G.A. Somorjai, Yimin Li, Proc. Natl. Acad. Sci. U.S.A. 2011, 108, 917-924. [2] B.J. Walder, C. Berk, W. Liao, ACS Cen. Sci. 2019, 5, 515-523. [3] D.B. Bucher, D.P.L. Aude Craik, M.P. Backlund, Nat.Protoc. 2019, 14, 2107-1747. [4] A.S. Ivanova. Kinet. Catal. 2012, 53, 425-439. [5] J.J Velasco-Vélez, V. Pfeifer. M. Hävecker. 2016, Rev. Sci. Instrum. , 87, 053121 INVITED AND PROMOTED LECTURES Acknowledgements: This study was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - 412351169 within the Emmy Noether program. R.R. acknowledges POSTERS support from the DFG Walter Benjamin Programme (project RI 3319/1-1). AH acknowledges funding from the European Unions Horizon 2020 research and innovation programme under ADVERTISERS the Marie Skodowska-Curie grant agreement No 841556. I.D.S. acknowledges support by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germanys AUTHOR INDEX Excellence Strategy EXC 2089/1 390776260. PO109 POSTERS / METHODS DEVELOPMENT 262 13 MEGHA MOHAN1, C NMR OF PARAMAGNETIC LAYERED DOUBLE ANDERS BRUHN ARNDAL HYDROXIDES (LDH) ANDERSEN2, JIRI MARES3 , Layered double hydroxides (LDH) find widespread application as catalysts and sorbents. They are layered inorganic ULLA GRO NIELSEN2, materials consisting of positively charged metal cation layers separated by an interlayer, which contain anions for JUHA VAARA1 charge neutrality and water [1]. LDH are notorious for structural disorder and lower crystallinity, which renders 1NMR Research Unit, University of Oulu, their characterization challenging. A series Mg Al-LDH doped with 0, 18%, 34%, 68% and 84% Ni(II) [2] with 13C 2 Oulu, Finland 2 labelled carbonate in the interlayer has been synthesized. 13C MAS NMR spectra of these materials has been record-Department of Physics, Chemistry, and Pharmacy, University of Southern ed. It was observed that the 13C NMR shielding tensor has three distinct eigenvalues, i.e, it possesses both anisotropy Denmark, and rhombicity (asymmetry) for all samples, but both the anisotropy and asymmetry are strongly dependent on the Odense, Denmark 3 concentration of the paramagnetic Ni ion doping. In contrast, axial symmetry is prediced based on the commonly Research Unit of Medical Imaging, Physics and Technology, University of used hydrotalcite crystal structure. Thus, we use computational approaches to explain the experimental observa- Oulu, Oulu, Finland tions and gain insight into the atomic level structure of LDH. The contributions to the total 13C shielding tensor of the intercalated CO 2- can be divided into diamagnetic con-3 tribution within the CO 2- ion and long-range paramagnetic contribution of the metal-oxygen layers. Since the an-3 isotropy and asymmetry are strongly dependent on the concentration of the Ni ions, we concentrate mainly on the long-range paramagnetic contribution due to dipolar part of the hyperfine coupling, modelled as a lattice sum of the electron-spin susceptibility a localized point dipole of the Ni centre [3]. This effect constitutes an important contribution to both the anisotropy and asymmetry, whereas the isotropic shift hardly changes as observed experi- INTRODUCTION mentally. Still, the experimental shift asymmetry is not quantitatively reproduced by the most precise single crystal SPONSORS structure (1M quintinite). On the other hand, other contributions to the anisotropy and asymmetry may arise for PROGRAM admixture of further structural polytypes (stacking disorder) and consideration of the hydrogen bonding situation PROGRAM — BY DAY of the carbonate anion. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] C. Forano, T. Hibino, F. Leroux, C. T-G. Ho, LAYERED DOUBLE HYDROXIDES. 2006, 1,1021-1092 [2] N.D. Jensen, C. Forano, S.S. Pushparaj, Y. ADVERTISERS Nishiyama, B. Bekele, U. G. Nielsen Phys. Chem. Chem. Phys. , 2018,20, 25335-25342 [3] L. Benda, J. Mares, E. Ravera, G. Parigi, C. Luchinat, M. Kaupp, J. Vaara, AUTHOR INDEX Angew. Chem., Int. Ed. , 2016, 55 , 14713. PO110 POSTERS / METHODS DEVELOPMENT 263 AKOKA SERGE, MISSTEC SEQUENCE, A NEW APPROACH FOR RF RENOU SOPHIE PULSES CALIBRATION Université de Nantes, CNRS, CEISAM UMR 6230, Nantes, NMR sequences are composed of multiple radio-frequency pulses. The probe adjustment, the sample concentra- France tion and the deuterated solvent influence the duration and the power of these pulses, impacting the applied angle. Therefore, it is important to calibrate them to reduce imperfections, loss of SNR, and to keep coherence selection. The commonly used method is to measure the nutation curve, variating the pulse duration or the pulse power to know the 90° angle. However, this method is impacted by the off-resonance effects, radiation damping, B and B in-1 0 homogeneities (shims) [1] even if an optimised model is used to fit this curve [2]. Furthermore, taking into account relaxation implies to know each T value. Most of the time these values cause a long acquisition time (up to 30 min 1 to obtain a complete and well digitized nutation curve). The MISSTEC sequence (alpha-(TE/2)-2*alpha-(TE/2)-acq-TM-alpha-TE/2-acq) was first proposed for mapping B in- 1 homogeneity in MRI [3], and later it was applied on high resolution spectrometers for the same purpose [4]. The flip angle alpha is obtained from the ratio between the stimulated and the spin echo which are sampled in presence of field gradient. Therefore, the results are not impacted by the B and B inhomogeneities or the relaxation times (T 1 0 1 and T ). Measurements performed with the MISSTEC sequence are comparable to nutation results for 1H. We show 2 that with this sequence it can be possible to have 0.3% of precision in 8 seconds for 1H. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] P.A. Keifer, Concepts Magn. Reson. 1999, 11, 165-180. [2] K. Nagashima, Concepts Magn. Reson. Part B (Magn. Reson. Eng.). 2012, 41, 1-12. [3] S. ADVERTISERS Akoka, F. Franconi, F. Seguin, A. Le Pape, Magneric Resonance Imaging. 1993, 11, 437-441. [4] A. Jerschow, G. Bodenhausen, J. Magn. Reson. 1999, 137, 108-115. AUTHOR INDEX Acknowledgements: The autors are grateful to Julien Pontabry and Gaëtan Assemat from RS2D Company for technical and scientific support. PO111 POSTERS / METHODS DEVELOPMENT 264 INCE RIDVAN1, THE MAGNETIC SPY: PROBING LOCAL MAGNETIC KERVERN GWENDAL1, PROPERTIES WITH SOLID-STATE NMR NICOLAS CLAISER1, THOMAS MAZET2 Acquisition of information on the magnetic properties at the molecular scale — namely the local magnetic susceptibility 1Université de Lorraine, — is important because it will lead to faster developments of advanced technological applications based on local magnetic Cristallographie, Résonance properties such as new methods for information storage [1], contrast agents for MRI [2], spintronics [3], etc… Magnétique et Modélisation, Vandœuvre-lès-Nancy, France 2 Polarized Neutron Diffraction [4], SQUID-based magnetometry [5], muon spin rotation [6], Electron Paramagnetic Reso-Université de Lorraine, Institut Jean Lamour, Nancy, France nance [7] give a precise insight on local magnetic susceptibility. These methods however require heavy equipment as well as relatively large crystalline samples which may be difficult to obtain. We implemented a model that has been tested and proven efficient in predicting paramagnetic SS-NMR spectra of microcrystalline powders [8]. This model calculates the effect of the hyperfine interaction on the NMR spectra. To this end, we need the crystallographic structure of the molecule. With the structure, the program will generate the position of all paramagnetic atoms on a given radius (50 Å) and set an initial rank-2 magnetic susceptibility tensor. This tensor is the only free parameter of this model and it affects the shape of the theoretical spectrum. From there, the program calculates the hyperfine interaction between all the paramagnetic atoms and one NMR observable nucleus and repeats this for each observable nucleus in the asymmetric unit cell. In the end, the program uses an optimization function to find the best agreement between the experimental and theoretical spectra, giving us an orientation of the local magnetic susceptibility tensor. INTRODUCTION This model was compared to experimental data on a series of isostructural lanthanide oxalate metal-organic SPONSORS frameworks. We will present and discuss our results on lanthanum, praseodymium and cerium oxalates. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] T. Matsumoto et al., Nat. Commun. 2014, 5, 3865. [2] Na, H. B. et al., Adv. Mater. 2009, 21, 2133–2148. [3] Manyala, N. et al., Nat. Mater. 2004, ADVERTISERS 3, 255–262. [4] Ridier, K. et al., Chem. - Eur. J. 2016, 22, 724–735. [5] Moro, F. et al., J. Magn. Magn. Mater. 2014, 2014, 361, 188–196. [6] Gygax, F. N. et al., Phys. B AUTHOR INDEX Condens. Matter 2003, 326, 359–363. [7] Žilić, D. et al., J. Magn. Reson. 2010, 207, 34–41. [8] Kervern, G. et al., Angew. Chem. 2009, 121, 3128–3132. PO112 POSTERS / METHODS DEVELOPMENT 265 ALEXANDRA SHCHUKINA1, ACQUISITION DURING TEMPERATURE CHANGES: SIGNAL P. MAŁECKI2, PROCESSING METHODS AND A PROTEIN NMR EXAMPLE B. MATEOS3, M. NOWAKOWSKI1, In protein NMR, temperature coefficients (TC), i.e., the dependen- 1) time-resolved NUS [1]: the data is divided into overlapping sub- M. URBAŃCZYK2, cies of chemical shifts on temperature, reveal valuable information. sets and reconstructed. This gives a boost in resolution/acquisi- G. KONTAXIS3, P. KASPRZAK2,4, For instance, non-linearities in TC indicate the presence of a protein tion time balance. The reconstruction we use is based on com- C. CONRAD-BILLROTH3, compact state. pressed sensing principle: out of all possible spectra, the spars- R. KONRAT3, est one is iteratively selected. K. KAZIMIERCZUK2 The acquisition for TC experiments, however, is hampered. One 2) a variant of the Radon transform [2], where temperature is treat-would need a series of at least 3-dimensional spectra. The acquisi- 1Faculty of Chemistry, Biological and ed as an additional dimension. Chemical Research Centre, University of tion of a single spectrum alone takes hours. The series should con- Warsaw, Poland tain a number of spectra with temperature increment. This number We applied the aforementioned approaches to temperature-swept 2Centre of New Technologies, University of Warsaw, Poland should be big enough to provide good resolution. Such acquisition 3-dimensional HNCO spectra of two intrinsically disordered pro-3Department of Structural and Computational will take too long to be practically feasible. teins - osteopontin and CD44 cytoplasmic tail [3]. Then, we estab- Biology, University of Vienna, Austria lished the non-linearities in their TC. The results were in line with 4Faculty of Physics, University of Warsaw, Here, we present two methods to overcome this obstacle. Both are Poland general biochemical considerations. based on non-uniform sampling (NUS): some points of the indirect dimensions are skipped and later reconstructed based on definite During my presentation, I will focus on the signal processing as-mathematical assumptions about the spectrum. One and the same pects of the two methods and show how they are complementary in NUS data-set can then be treated by both methods. terms of sensitivity and resolution. INTRODUCTION The software developed in our group for both processing methods is SPONSORS available at www.nmr.cent.uw.edu.pl → Downloads. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] Mayzel, M., Rosenlöw, J., Isaksson, L., Orekhov, V. Y. J. Biomol. NMR 2014, 58, 129 [2] Dass, R., Kasprzak, P., Kazimierczuk, K. J. Magn. Reson. 2017, 282, 114 [3] A. Shchukina, P. ADVERTISERS Małecki, B. Mateos, M. Nowakowski, M. Urbańczyk, G. Kontaxis, P. Kasprzak, C. Conrad-Billroth, R. Konrat, K. Kazimierczuk, Chem. Eur. J. 2021, 27, 1753. AUTHOR INDEX Acknowledgements: This work was supported by National Science Centre, Poland, grant HARMONIA 501-D313-0000174. PO113 POSTERS / METHODS DEVELOPMENT 266 MATEUSZ A. SŁOWIŃSKI, ROTATIONAL DIFFUSION OF CHIRAL MOLECULES WŁODZIMIERZ MAKULSKI, IN THE ELECTRIC FIELD STUDIED BY PIOTR GARBACZ University of Warsaw, Faculty of MOLECULAR DYNAMICS Chemistry, Warsaw, Poland The electric field E perturbs the rotational diffusion of a molecule that bears the permanent electric dipole moment. Studies of this perturbation are especially pronounced if one considers a chiral molecule. In this case, it has been predicted that interaction involving the so-called antisymmetric part of the indirect spin-spin coupling ( J*) generates signals which allow direct discrimination between enantiomers [1, 2], i.e. , without the usage of chiral derivatizing agents and solvents. This effect may be analyzed using tensor calculus, but in order to include the fact that the molecule is subjected to friction in the solution, i.e. , it cannot follow the time-varying E field instantly, one needs to use more advanced models of the rotation diffusion such as those based on the molecular dynamics. We chose two chiral compounds – L-alanine and ( R)-1,1,1-trifluoropropan-2-ol. In the former molecule, the spin system was 19F-1H, i.e. , C19F -HC(O1H)-CH ), and in the latter 15N-13C . In order to provide the most reliable data for 3 3 a our simulations of the predicted effect in trifluoropropanol, we used the spin-spin coupling determined from the gas-phase NMR studies. We found that extrapolated to the zero-density limit spin-spin coupling 3 J(19F, 1H) is (6.5 ± 0.5) Hz. This value agrees with the results of quantum chemical computations performed in Dalton. Moreover, the computations indicate that the antisymmetric parts of the studies couplings are: 1 J*(15N, 13C) = -0.332 Hz and 3 J*(19F, 1H) = 1.25 Hz. INTRODUCTION Molecular dynamics calculations have been carried out with the GROMACS program for the alanine zwitterion in SPONSORS water and trifluoropropanol dissolved in ethanol (simulation length > 10 ns, box size ~6×6×6 nm). We considered PROGRAM two cases: ( i) there is no electric field; thus, isotropic rotational diffusion occurs, and the effect is not present (es- PROGRAM — BY DAY timation of the uncertainty of the simulation), and ( ii) application of the E field of the amplitude 1 V/nm and fre- TOPICS quency 3 GHz (computation of the amplitude of the effect). We quantified the influence of the electric field on the PRIZE LECTURES rotational diffusion by evaluation of the ensemble-averaged products of Wigner functions. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] P. Garbacz, A. D. Buckingham, J. Chem. Phys. 2016, 145, 204201. [2] J. P. King, T. F. Sjolander, J. W. Blanchard, J. Phys. Chem. Lett. 2017, 8, 710-714. AUTHOR INDEX Acknowledgments: PG and MS acknowledge the National Science Centre, Poland, for the financial support through OPUS 16 Grant No. 2018/31/B/ST4/02570. PO114 POSTERS / METHODS DEVELOPMENT 267 ARTUR SOLODOVNYK, ELECTRICALLY DETECTED MAGNETIC RESONANCE OLEKSII LAGUTA, SETUP BASED ON A NOVEL THz EPR SPECTROMETER PETR NEUGEBAUER Central European Institute of Electrically detected magnetic resonance (EDMR) is a sensitive and powerful technique for the determination of Technology, Brno University of fundamental intrinsic properties of semiconductive solid-state materials. We develop an EDMR setup based on the Technology, Brno, Czech Republic terahertz EPR spectrometer located in CEITEC BUT, which will operate at frequencies up to 1.1 THz and external magnetic fields up to 16 T. In this way, our main THz EPR setup will allow scanning both the magnetic field and frequency. Thus, compared with the conventional CW EDMR, this will allow obtaining not only the dependence of the voltage change on the external magnetic field at a certain frequency, but also to take a frequency-field map [1]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] P. Neugebauer et al., Phys. Chem. Chem. Phys. 2018, 20, 15528-15534. ADVERTISERS Acknowledgements: This research was carried out under the project CEITEC 2020 (LQ1601) with financial support from the Ministry of Education, Youth and AUTHOR INDEX Sports of the Czech Republic under the National Sustainability Programme II. PO115 POSTERS / METHODS DEVELOPMENT 268 DEEPANSH SRIVASTAVA1, CORE SCIENTIFIC DATASET MODEL: THOMAS VOSEGAARD2, A LIGHTWEIGHT AND PORTABLE MODEL AND DOMINIQUE MASSIOT3, PHILIP GRANDINETTI1 FILE FORMAT FOR MULTI-DIMENSIONAL 1Department of Chemistry, Ohio State SCIENTIFIC DATA University, Columbus, OH, USA 2Laboratory for Biomolecular NMR Spectroscopy, Department of Molecular With increasing pressure from the funding agencies and scientific journals to archive and share primary and pro-and Structural Biology, University of cessed data, there is a growing sense of urgency for a stable, resourceful, and future-proof file format for the ex-Aarhus, Aarhus C, Denmark 3 change of scientific datasets. We address this problem by proposing a Core Scientific Dataset (CSD) Model[1] that CEMHTI UPR3079 CNRS, Univ. Orléans, Orléans, France can encode a wide variety of multi-dimensional and correlated datasets. The CSD model with JavaScript Object Notation (JSON) serialization is presented as a lightweight, portable, and versatile standard for intra- and interdisciplinary scientific data exchange. This model supports datasets with a p-component dependent variable, {U ,… 0 ,U , …,U }, discretely sampled at M unique points in a d-dimensional independent variable (X ,…X ,…X ) space. q p−1 0 k d−1 Moreover, this sampling is over an orthogonal grid, regular or rectilinear, where the principal coordinate axes of the grid are the independent variables. It can also hold correlated datasets assuming the different physical quantities (dependent variables) are sampled on the same orthogonal grid of independent variables. The model encapsulates the dependent variables’ sampled data values and the minimum metadata needed to accurately represent this data in an appropriate coordinate system of independent variables. The CSD model can serve as a reusable building block in the development of more sophisticated portable scientific dataset file standards. It is currently supported INTRODUCTION by the csdmpy Python package, SIMPSON, EasyNMR, DMFit, and RMN. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] D. Srivastava, T. Vosegaard, D. Massiot, and P.J. Grandinetti, PLOS ONE 2020, e0225953. PO116 POSTERS / METHODS DEVELOPMENT 269 MATÚŠ ŠEDIVÝ1, SOFTWARE FOR VERSATILE HFEPR SPECTROMETER ANTONIN SOJKA2, VINICIUS SANTANA2, Even though nowadays some High Frequency Electron Paramag- and FDMR measurements it is possible to use a scripting feature to ANDREJ GABRIŠ3, netic Resonance (HFEPR) spectrometers are commercially avail- automatically execute a sequence of measurements. An integrated OLEKSI LAGUTA2, able, most of them are still custom-built machines. A versatile script editor utility can generate a batch of measurement sequence PETR NEUGEBAUER2 HFEPR spectrometer with a Frequency Rapid Scanning (FRaScan) with sweep of desired parameter such as microwave frequency or 1 feature was built at CEITEC BUT [1]. A specific software solution for magnetic field, orientation and temperature. Because FDMR mea-Brno University of Technology, Department of Microelectronics, control of spectrometer was developed in LabVIEW [2] because it surements are much faster to perform and can yield a large data, Brno, Czech Republic simplifies memory management, parallel execution, and commu- the TDMS file format was used to organise datasets obtained from 2Brno University of Technology, nication with instruments via standard interfaces. The software is whole measurement sequence. The Zeeman diagram measure-Central Europen Institute of Technology, Brno, Czech Republic composed of individual modules for each instrumental part and ments are done by performing an FDMR measurements during con-2Brno University of Technology, main controller, which are connected via messaging service. This tinuous sweep of a magnetic field Faculty of Mechanical Engineering, Brno, Czech Republic allows modules to run independently on each other, distribute tasks To demonstrate capabilities of spectrometer and its automated con- in threads and decrease average CPU load. The main controller is trol we performed a measurements on single crystal of copper ace-the top software component that provides an event driven user in- tate as a model system for molecular magnetism. Measurements in terface, as well as execution of a measurement routine and distribu- a frequency range 205-245 GHz, at temperature 200 K and magnetic tion of messages to modules. field 7.5 T were done in less than 2 hours for 280 orientations with Currently, there are 3 types of user interface for different tasks. One step 1 degree. For comparison, alike measurements in a CW mode for Continuous Wave (CW) measurements, second for Frequen-would take approximately 30 hours. Obtained data were post pro- INTRODUCTION cy Domain Magnetic Resonance (FDMR) measurements and third cessed in orientation vs frequency map. Additionally, a Zeeman dia- SPONSORS for Zeeman diagram measurements [3]. All of them have a similar gram was measured in a field range 4.5 to 9 T, and a frequency range PROGRAM design and share most of the back-end components. In case of CW 180 to 240 GHz. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] A. Sojka, M. Šedivý, O. Laguta, Electron Paramagnetic Reonance 2020, 27, 214-252. [2] C.J. Kalkman, J Clin Monitor Comput. 1995, 11, 51-58. [3] P. Neugebauer, D. Bloss, R. Marx, POSTERS Phys. Chem. Chem. Phys. 2018, 20, 15528-15534. ADVERTISERS Acknowledgements: This work was financially supported from the ERC under the European Unionś Horizon 2020 research and innovation programme (GA No 714850) and BUT internal AUTHOR INDEX agency grant FEKT-S-20-6215. PO117 POSTERS / BENCHTOP AND LOW-FIELD NMR 270 ANTON DUCHOWNY1, COMPACT HIGH-PRESSURE GAS-PHASE PABLO MATIAS DUPUY2, NMR SPECTROSCOPY HEGE CHRISTIN WIDERØE2, OLE JOHAN BERG2, A new, low-cost, and highly versatile setup for benchtop NMR spectroscopy and relaxation measurements of gases AUDUN FAANES2, at high-pressure is introduced [1]. It utilizes mostly commercial parts, which can be easily exchanged, and it in-ANFINN PAULSEN3, cludes multiple safety features. Except for the gas bottles, the novel high-pressure (HP) setup is small enough to fit HOLGER THERN4, into a 1.2 m wide fume hood. It enables pressures up to 200 bar on single gas components and mixtures of multiples OLIVER MOHNKE4, gases. A chosen pressure can be set with the help of a mixing chamber within a couple of seconds. MARKUS KÜPPERS1, BERNHARD BLÜMICH1, The versatility of the setup is demonstrated in the first step by quantitatively analyzing the pressure dependence ALINA ADAMS1 of the pure methane and the composition of a gas mixture containing two hydrocarbons and hydrogen at 200 bar 1 total pressure with the help of 1H NMR spectra. The sensitivity of modern benchtop NMR spectrometers is so good Institut für Technische und Makromolekulare Chemie, RWTH Aachen that a signal-to-noise-ratio above 104 can be achieved, although densites of hydrocarbon gas mixtures are about 6 University, times lower compared to water. Moreover, the spectral resolution of modern benchtop NMR spectrometers is high Aachen, Germany 2 enough to quantify the gas composition of this particular self-made 3-component mixture simply by integration of Equinor ASA, Ranheim, Norway 3Gassco AS, Kopervik, Norway the signals of interest without the need of a spectral deconvolution software. 4Baker Hughes INTEQ GmbH, Celle, Germany Furthermore, the newly designed HP setup was applied to investigate a gas-solid interaction by exposing solid poly-vinyl chloride (PVC) to CO gas. Although CO is invisible in proton NMR, the ingress of gaseous and eventually 2 2 supercritical CO into the polymer matrix can be observed by changes in the PVC spectrum in terms of 1H linewidth 2 INTRODUCTION and peak integral. In addition the gas-solid interaction, the gas-liquid interaction between benzene and methane is SPONSORS observed. As methane pressure increases, the mole fraction of dissolved gas rises accordingly. PROGRAM The presented results demonstrate that the proposed HP-setup bears great promises in various gas applications. We PROGRAM — BY DAY expect that this novel setup will open up new opportunities also for investigations which currenly largely rely on TOPICS other analytical methods, such as gas chromatography. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] A. Duchowny et al., Journal of Magnetic Resonance; submitted ADVERTISERS Acknowledgements: We acknowledge the financial support from Equinor, Gassco and Baker-Hughes. Also, we would like to thank our colleagues at the ITMC AUTHOR INDEX mechanical workshop and the scientists from Magritek for the help designing and building the hardware. PO118 POSTERS / BENCHTOP AND LOW-FIELD NMR 271 TUGBA DISPINAR GEZER1, LOW FIELD NMR CHARACTERIZATION OF TISSUE MIMICKING ILKER UN1, GELS FOR MRI PHANTOM APPLICATIONS ADRIANO TROIA2, GOKHAN AKTAS1 Magnetic Resonance Imaging (MRI) phantoms are calibration ob- they can be shaped in more more complex geometries, serving the 1The Scientific & Technological Research jects which provide tissue-equivalent properties (e.g. relaxation production of heterogeneous phantoms in the shapes and sizes of Council of Turkey, National Metrology times, electrical conductivity, etc), used to evaluate the effectiveness human organs, without the need for rigid containers and septa. Institute, TUBITAK UME, Turkey 2Istituto Nazionale di Ricerca and performance of MRI scanners and quantitative MR algorithms. Therefore, gels are recognized as interesting materials in the devel-Metrologica (INRiM), Turin, Italy An MRI phantom, as a standard system, should exhibit anatom- opment of MRI phantoms. Especially, polysaccharides such as aga- ical, physiological, or bio/chemical characteristics that are do not rose, agar, gelatin, carrageenan, and synthetic polymers such as pol-change over time, and result to be comparable between scanners, yvinyl alcohol, polyacrylamide are well-known materials to prepare manufacturers, and measurement protocols. Therefore, an MRI the MRI phantoms because of their physically crosslinkable feature phantom should have traceable, validated, and monitored compo- [2, 3]. These materials dissolve in hot water and upon cooling turn nents. In this sense, the characterization of phantom materials with out to a semi-solid gel, giving the advantage of easy preparation. respect to physical Nuclear Magnetic Resonance (NMR) properties In this work, gellan gum, and agar based polysaccharide gels have and stability is essential [1]. Especially, low field NMR systems, with been studied in order to mimic the relaxation properties of brain their magnetic field strengths corresponding to those of MRI scan-tissues (white matter, grey matter) and cardiac muscle. Gellan gum ners (1.5 T, 3 T) are valuable reference platforms to characterize the has been chosen as the gel matrix to increase the strength and the relaxation times of phantom materials and to check their long-term mechanical stability of the gel while different amounts of GdCl and stability. 3 Agar have been used in the formulation to tune T (spin-lattice), T 1 2 INTRODUCTION MRI phantoms, typically composed of paramagnetic ions such as (spin-spin) relaxation times, respectively. NaN has also been added 3 SPONSORS CuSO , NiCl , MnCl , or GdCl in order to have comparable relaxa- as a preservative to retard mold formation over time. 1.4 T (60 MHz) 4 2 2 3 PROGRAM tion times to those of human tissues, are prepared either as aqueous desktop NMR machine has been used in the characterization of T , 1 PROGRAM — BY DAY solutions or as gels. Upon two, gels give the advantage of making T relaxation times of the gels as well as for monitoring their long-2 TOPICS large and strong phantoms. Moreover, being robustly processable, term stability. PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] K. E. Keenan, et al., Magn Reson Med 2018, 79, 48-61. [2] K. Yoshimura, Magn Reson Med 2003, 50, 1011-1017. [3] K.J.M. Surry, et al., Phys. Med. Biol. 2004, 5529-5546. ADVERTISERS Acknowledgements: The results here presented have been developed in the framework of the EMPIR Project 18HLT05 QUIERO. This project has received funding from the EMPIR programme AUTHOR INDEX co-financed by the Participating States and from the European Union’s Horizon 2020 research and innovation programme. PO119 POSTERS / BENCHTOP AND LOW-FIELD NMR 272 ANTON DUCHOWNY AND NOVEL LOW-COST NMR METHOD FOR IDENTIFICATION AND ALINA ADAMS QUANTIFICATION OF PVC PLASTICIZERS Institut für Technische und Makromolekulare Chemie, RWTH Polyvinyl chloride (PVC), the third most produced polymer word- and quantification of the various plasticizers can be easily done us- Aachen University, Aachen, Germany wide and has a large variety of applications ranging from medical ing characteristic signals in a spectral range above 2.5 ppm and in the to industrial products. To meet the largely different property represence of non-deuterated solvents which have signals outside the quirements, various plasticizers are added in different amounts to spectral range of interest. For this purpose, non-deuterated n-hex-the PVC product. Yet, in most cases, the plasticizers are only mixed ane, a suitable solvent for PVC, was chosen as an alternative to more with the PVC matrix and consequently they tend to migrate out of costly deuterated solvents. The use of appropriate non-deuterated the PVC product with time. This leads to a deterioration of the PVC solvents, shown here for the first time, is very appealing due to their product properties as the amount and type of plasticizer have a crit-much lower costs compared to the costs of the deuterated solvents, ical impact on them. Moreover, legal regulations exist about the use which are the standard solvents used for liquid-state NMR spectros- of particular plasticizers in contact with human skin and groceries. copy. The reliability of the proposed method using non-deuterated Thus, it is of key importance to identify high throughput analytical solvents is demonstrated by comparisons with the spectra recorded methods which are able to identify the used plasticizers and quan-on the same plasticizer dissolved in deuterated chloroform solvent. tify their amounts. To meet these current challenges, our work in- Given that low-field NMR equipment is, by far, more affordable then troduces a new, simple, and low-cost experimental methodology for the high-field NMR and the costs for maintenance, extra personnel, the identification and quantification of PVC plasticizers in solution and facilities are negligible, the shown results indicate that the pro-or in a product [1]. It is based on low-field 1H spectra acquired with a posed low-field NMR analysis is a low-cost alternative for the study INTRODUCTION low-cost and easy to operate NMR benchtop device and the usage of of PVC plasticizers. Plasticizer concentrations below 2 mg mL−1 in SPONSORS appropriate non-deuterated solvents. solution, corresponding to 3 wt% in a PVC product, can be quanti- PROGRAM Despite being rather complex molecules with molar masses around fied within only 1 minute. Due to its simplicity, the proposed meth- PROGRAM — BY DAY 200 g mol-1- 500 g mol-1, the 1H spectra of all investigated plasticiz- odology is appealing even to non-NMR experts and can be applied TOPICS ers show well separated peak-clusters for aromatic (~7 ppm), a-CH to identify and quantify plasticizers of large quantities of polymer 2 PRIZE LECTURES (~4 ppm) and aliphatic groups (1-2 ppm). Since the aliphatic peak samples in a short experimental time. PLENARY LECTURES regions of various plasticizers are rather similar, the discrimination TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] A. Duchowny, A. Adams, Molecules 2021, 26(5), 1221. PO120 POSTERS / BENCHTOP AND LOW-FIELD NMR 273 GOSSUIN YVES1, FOLLOWING CHROMIUM (IV) REDUCTION BY LOW RESOLUTION SOPHIE LAURENT2, NUCLEAR MAGNETIC RELAXOMETRY DUEZ PIERRE3, MASSON CÉLINE1, Chromium is released in the effluents of several industries. It is for At higher concentrations of chromium and in complex systems, BLANKERT BERTRAND4 example used for stainless steel production, leather tanning and nuclear magnetic resonance relaxometry, based on the magnetic 1University of Mons UMONS, electroplating. Cr(VI), mostly of anthropogenic origin, is very solu- properties of chromium, could be applied. Indeed, all the orbitals Biomedical Physics Unit, Mons, ble and toxic for the flora and fauna, even at low concentration in of Cr(VI) are full which results in a null magnetic moment. On the Belgium 2University of Mons UMONS, water (0.1 mg/L) while Cr(III) is an essential nutrient (daily intake of contrary, the electronic configuration of Cr(III) (3s23p63d3) results in General, Organic and Biomedical 25 - 35 µg for an adult) that is only toxic at high concentrations1. One a non-null magnetic moment of 3.97 µ . Cr(III) is thus paramagnetic B Chemistry Unit, Mons, Belgium 3 strategy to reduce the toxicity of chromium in water is thus the re- and will shorten the water proton relaxation times T and T in aque- University of Mons UMONS, 1 2 Therapeutic Chemistry duction of noxious Cr(VI) to Cr(III). Several compounds can be used ous solutions7. In this study we show that the reduction of Cr(VI) to and Pharmacognosy Unit, Mons, to achieve this reduction such as hydrogen peroxide2, Fe(II) ions3, Cr(III) by hydrogen peroxide in acidic conditions can be followed Belgium 4University of Mons UMONS, zerovalent aluminium4 and plant extracts5. The monitoring of the through the shortening of water T and T . The relaxation rates 1/ T 1 2 1 Laboratory of Pharmaceutical Analysis, reduction is often carried out by UV-Visible spectrometry after the and 1/ T of partially reduced Cr(VI) solutions are linearly depending 2 Mons, Belgium addition of diphenylcarbazide to the solution6, the so-called “carba- on the actual Cr(III) content of the mixture, which was determined zide test” . This technique is really sensitive since it allows the detec- using direct UV-visible spectroscopy, without the use of carbazide8. tion of Cr(VI) in water even at very low concentrations (0.002 mg/L). In order to determine the best experimental conditions, the field, However, it is destructive for the sample and can only be used for temperature and pH dependences of the relaxation rates of com-clear solutions which is a limitation when using reductants as pow- pletely reduced solutions have been studied. The use of T seems 2 INTRODUCTION ders of zerovalent aluminium or iron, and when working directly more suited to follow the chromium reduction, since it is faster to SPONSORS on wastewater. Moreover, the quantification with carbazide must be measure than T and also because 1/ T increases with the field while 1 2 PROGRAM achieved in a given concentration range in order to remain linear 1/ T decreases. 1 PROGRAM — BY DAY and cannot be used at Cr(VI) concentrations larger than 2 mg/L. TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] P.B. Tchounwou et al, EXS. 2012, 101, 133–164. [2] M. Pettine et al, Environ. Sci. Technol. 2002, 36, 901-907. [3] I.J. Buerge, S.J. Hug, Environ. Sci. Technol. 1997, 31, 1426-1432. [4] POSTERS C.J. Lin et al, Water Res. 2009, 43, 5015-5022. [5] V.A. Okello et al, Environ. Sci. Technol. 2012, 46, 10743-10751. [6] R.T. Pflaum, L.C. Howick, J. Am. Chem. Soc. 1956, 78, 4862-4866. [7] I. Bertini ADVERTISERS et al, Inorg. Chem. 2001, 40, 4030-4035. [8] A. Sanchez-Hachair, A. Hofmann, C. R. Chimie. 2018, 21, 890-896. AUTHOR INDEX Acknowledgements: F.R.S.-FNRS for the financial support. PO121 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 274 NARGIZ ASANBAEVA1, 2, EPR STUDY OF HIGHLY STABLE BIRADICALS DENIS MOROZOV1, PERSPECTIVE FOR DNP IN CELL SERGEY DOBRYNIN1, OLGA ROGOZHNIKOVA1, Dynamic Nuclear Polarization (DNP) is a powerful technique to improve the sensitivity of solid state NMR exper- DMITRY TRUKHIN1, iments on biological materials, which allows high-resolution spectra to be obtained. Biradicals are often used as IGOR KIRILYUK1, polarizing agents due to the relatively strong exchange interaction between the paramagnetic moieties[1]. But re-VICTOR TORMYSHEV1, cent studies showed the crucial factor for CE-DNP is not the large sum, spin exchange interaction ( J) and dipolar ELENA BAGRYANSKAYA1 coupling ( D), but rather the relative magnitude of J and D, expressed as the J/ D ratio [2]. Therefore, the problem 1 Novosibirsk Institute of Organic Chemistry, arises of obtaining biradicals with a certain value of spin exchange. Also when working with polarizing agents, Novosibirsk, Russia 2 there are problems with their solubility in water and their stability in reducing media which is very important at the Novosibirsk State University, Physics Department, Novosibirsk, Russia biological sample preparation. If the problem with water solubility in some biradicals is solved, but their stability is uncertain. According to previous studies the stability of nitroxides depends on ring size and substituents adjacent to the paramagnetic center [3]. In this work we studied the biradicals based on five-membered ring nitroxides with tetraethyl substituents, which will provide additional stability. Also, the use of trityl as one of the paramagnetic centers makes it possible to increase the resistance to reduce and attenuate the effect of depolarization at high magnetic field [4]. A series highly stable nitroxyl-nitroxyl and trityl-nitroxyl with different J values were synthesized and their magnetic resonance parameters as well as stability against reduction by ascorbic acid in buffer solution at pH 7.4 were studied. The stability of obtained DNP agents is very high and can allows to use them in the cell. One biradical based on low toxic INTRODUCTION OX063 trityl and tetraethyl pyrrolidine radical reveal the highest stability and water solubility. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] F. Mentink-Vigier et al, J. Am. Chem. Soc. 2018, 140, 11013-11019. [2] A. Equbal, K. Tagami, S. Han, Phys. Chem. Chem. Phys. 2020, 22, 13569– ADVERTISERS 13579. [3] I. Kirilyuk et al, J. Org. Chem. 2015, 80, 9118–9125. [4] F. Mentink-Vigier et al, Chem. Sci. 2017, 8, 8150–8163. AUTHOR INDEX Acknowledgements: The work was supported by the Ministry of Science and Higher Education of Russia (project number 14.W03.31.0034). PO122 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 275 XIAOXUN CHEN, X-BAND EPR STUDY OF A POTENTIAL CHEMICAL SYSTEM FOR VENKATA SUBBARAO INDIRECT PHOTO-DNP REDROUTHU, HOLGER HILBERT, In certain organic molecules, triplet states are generated by pulsed solutions of toluene. They observed the transient triplet signal from SANJAY VINOD KUMAR, laser excitation of the sample. The excited triplet state is populat- the quinones as well as a strong emissive transient signal from the GUINEVERE MATHIES ed via intersystem crossing (ISC) from the excited singlet and of- nitroxides. The latter suggests that the nitroxides obtain a strong, Department of Chemistry, University of ten exhibits a non-Boltzmann polarization. It is possible to exploit non-thermal polarization. In our laboratory, we reproduced the re-Konstanz, Konstanz, Germany this high polarization and directly transfer it to nuclei to enhance sults of Tarasov et al. using an excimer laser as the photo-excitation the NMR signal in a direct photo-DNP, or triplet DNP, experiment. source. We expanded the transient EPR data set and simulated the In indirect photo-DNP, one transfers the polarization from triplet triplet spectra of the quinones using EasySpin.[2]. We found that all electrons first to a doublet species. The surrounding nuclei obtain three quinones show two triplet species with distinct ZFS parame-the polarization from the doublet species in a consecutive step. The ters. Furthermore, to confirm the enhanced nitroxide polarization, large majority of the photo-DNP experiments has been done with we recorded echo-detected EPR spectra at X band. Upon photo-ex-pentacene, and the polarization transfer was direct. But there are citation these spectra showed a decrease of the absorptive TEM-disadvantages to this molecule such as the poor photo-stability. PO signal, which could result from emissively polarized nitroxide Also, so far for all photo-DNP has been done at low field (X-band). radicals, but also from sample heating by the laser. However, when Therefore, we are exploring a new chemical system for indirect pho-performing an inversion recovery experiment, we observed an in- to-DNP, which could possibly also be used for high-field photo-DNP. crease of the inverted echo signal when the laser flash precedes the π-pulse. This suggests that at least some of the nitroxide radicals In 2002, Tarasov et al. reported non-equilibrium spin polarization INTRODUCTION in the sample have obtained an enhanced polarization. To improve of nitroxide radicals upon photo-excitation of molecular complexes SPONSORS our experiment, we are currently assembling a laser set up, which of a nitroxide radical and a quinone, which form in frozen toluene PROGRAM will enable us to generate triplets in our samples more efficiently. [1]. They measured the transient X-band (9.7 GHz) EPR spectra of PROGRAM — BY DAY a series of para-quinones with nitroxide radical (TEMPO) in frozen TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] V. F. Tarasov, I. A. Shkrod, A. D. Trifunac, J. Phys. Chem. A. 2002, 106, 4838-4845. [2] S. Stoll, A. Schweiger, J. Magn. Reson. 2006, 178(1), 42-55. PO123 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 276 SABRINA ELKHANOUFI, NEW, HIGHLY SENSITIVE OFF/ON EPR PROBES RACHELE STEFANIA, TO MONITOR ENZYMATIC ACTIVITY DIEGO ALBERTI, SIMONA BARONI, Many pathologic conditions are often associated with unregulated bond to yield Tempo-C12 (TC12) and Tempo-2-C12 (T2C12). Both SILVIO AIME, level of enzyme activity. Therefore, the detection and quantification compounds exhibit a low solubility in water and aggregate to form SIMONETTA GENINATTI CRICH of the enzymatic activity is extremely important for a diagnostic pur- stable micelles with the lipophilic tail in the core and the nitroxide University of Torino, Molecular Biotechnology pose. [1] In this contest, a particularly interesting class of enzymes radical exposed to water. The radicals in the micellar aggregates and Health Sciences, Torino, Italy is represented by carboxylesterases (CEs). These enzymes belong to are practically EPR silent showing a low and broad EPR signal. The the serine hydrolase superfamily and are involved in the hydrolysis hydrolysis of the ester bond catalyzed by CEs release two different of endogenous ester-containing substrates as well as ester-contain-nitroxide radicals: 4-Hydroxy-TEMPO and 4-Oxo-TEMPO. The first ing drugs, thus playing a crucial role in a variety of metabolic pro- one is released from the TC12 micelles while the second from T2C12 cesses. [2] CEs are upregulated in many tumors and the assessment micelles and they generate an intense and narrow EPR signal, that is of their activity may be of diagnostic interest as well it may provide proportional to the enzymatic activity. [4,5] relevant information regarding chemotherapeutic effects of anti- CEs1, CEs2 and esterase from porcine liver (PLE) were tested. The re- tumor ester-containing drugs and pro-drugs. Currently CE activity sult obtained show that the micelles of TC12 and T2C12 have a much is assessed by means of fluorescence and UV-based methods. [3] In higher selectivity toward the CEs2, and a Limit of Detection of the this study, we propose the use of electron paramagnetic resonance same order of those ones obtained with optical methods. In conclu- (EPR) as an easy method to probe CE enzymatic activity in vitro. EPR sion, this is a new promising tool to quantitatively detect the CEs2 has the advantage to be highly sensitive and with limited interfer- activity showing an interesting off/on EPR signal after the enzymat- INTRODUCTION ences from the matrix, also in the presence of turbid samples. ic activity. The method can be applied for monitoring the enzymatic SPONSORS For this application, TEMPO derivative nitroxide radicals were con- activity in vivo, eventually also through the detection of the Over- PROGRAM jugated to a fatty acid (Dodecanoic acid) via the formation of an ester hauser MRI response. PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES References: [1] Ou Y., Wilson R., Weber S., Annu. Rev. Anal. Chem. 2018, 11, 509–33. [2] Wang D., Zou L., Jin Q., Hou J., Ge G., Yang L., Acta Pharmaceutica Sinica B 2018, 8 (5), 699–712. [3] Lan POSTERS L., Ren X., Yang J., Liu D., Zhang C., Bioorg. Chem. 2020, 94, 103388. [4] Audran G., Jacoutot S., Jugniot N., Marque S., Mellet P., Anal. Chem. 2019, 91 (9), 5504-5507. [5] Aliaga C., Rezende M., ADVERTISERS Mena G., Magn., Reson. Chem. 2016, 54, 870–873. AUTHOR INDEX Acknowledgements: This project has received funding from the European Union's Horizon 2020 research and innovation program under grant agreement No 86309, Primogaia project. PO124 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 277 BURKHARD ENDEWARD1, LONG RANGE DISTANCE DETERMINATION ON YANPING HU2, FULLY DEUTERATED RNA XIANYANG FANG2, THOMAS PRISNER1 PELDOR (pulsed electron electron double resonance) also called DEER (double electron electron resonance) is a 1Institute of Physical and Theoretical magnetic resonance method for to determine the distance and the distance distribution in double spin-labeled Chemistry and Center for Biomolecular macromolecules like proteins, RNA, or DNA as well as polymers [1,2]. The maximum accessible distance r is lim-Magnetic Resonance (BMRZ), Goethe max University, Frankfurt, Germany ited by the dipolar observation time window t spanned by the pulse settings [3]. This dip 2Beijing Advanced Innovation Center time window depends strongly on the phase memory time of the nitroxide spin labels. The phase memory time for Structural Biology, School of Life at the experimental temperature of 50 K is manly determined by the coupling to other spins in the sample and Sciences, Tsinghua University, Beijing, China depends on the gyromagnetic ratio of such spins. Therfore protons and other electrons are the main contributors. Basic techniques to get rid of such spins is substituting protons by deuterons and reducing the radical (and therefore sample) concentration [4,5]. In this contribution we demonstrate that PELDOR experiments on fully deutereated RNA molecules allow to adress distances up to 10 nm quantitatively and to analyse conformational flexibility of long RNAs. We demonstrate this on a fully deuterated dengue virus (DENV) 3’SL RNA construct [6,7], using an unnatural base pair in a modified genetic code for posttranscriptional spin labelling [8]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] A. Milov, K. Salikov, M. Shirov, Fiz. Tverd. Tela. , 1981, 23, 975-982. [2] M. Pannier, S. Veit, A. Godt, G. Jeschke, and H. W. Spiess, J. Magn. Reson.. 2000, 142, 331-340. [3] Jeschke, G. Annu. Rev. Phys. Chem. 2012, 63, 419–446. [4] R. Ward, A. Bowman, E. Sozudogru, H. El-Mkami, T. Owen-Hughes, and INVITED AND PROMOTED LECTURES D. G. Norman, J. Magn. Reson.. 2010, 207, 164-167. [5] [1]T. Schmidt, M. A. Wälti, J. L. Baber, E. J. Hustedt, and G. M. Clore, Angew. Chem. Int. Ed. . 2016, 55, POSTERS 15905–15909. [6] Y. Wang, Y. Chen, Y. Hu, and X. Fang, PNAS. 2020, 117, 22823–22832. [7] Y. Zhang et al., EMBO Rep. , 2019, 20(11), e47016. [8] Y. Wang et al., ADVERTISERS Chem. Sci. 2020, 11, 9655-9664. AUTHOR INDEX Acknowledgements: Sino-German Center for Research Promotion (SGC) Fund. PO125 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 278 AATHIRA GOPINATH AND THE OUTER MEMBRANE INSERTASE BAMA EXCURSES OVER A BENESH JOSEPH BROAD CONFORMATIONAL SPACE IN NATIVE ENVIRONMENT Institute of Biophysics, Department of Physics and Centre for Biomolecular Outer membrane proteins (OMPs) are involved in several cellular the unique properties of the outer membrane in vitro, which largely Magnetic Resonance, Goethe University, Frankfurt am Main, Germany processes vital for the survival of Gram-negative bacteria. The prop- hindered a thorough understanding of the BAM function. er folding and insertion of the unfolded OMPs into the outer mem- Here we used in situ pulsed electron-electron double resonance brane is a crucially regulated process. In E. coli, this process is cata- spectroscopy (PELDOR or DEER [3]) to investigate the conforma- lysed by the β-barrel assembly machinery (BAM) complex. BAM is tional states of BamA in the native membrane or in the LDAO de-a heterooligomeric complex composed of the central β-barrel pro- tergent micelles. Cysteine pairs were introduced at the extracellular tein BamA and its associated lipoproteins BamB-E. BamA is high- and periplasmic areas of BamA near the lateral gate. These cysteine ly conserved and essential, making it an attractive target for novel pairs were labeled with MTSL either in the LDAO micelles or directly antibiotics. In BamA, the C-terminal transmembrane domain is in the isolated native outer membranes. We experimentally deter-connected to five N-terminal polypeptide transport associated (PO- mined the distances between the extracellular loops L1-L8, L3-L8, TRA) domains located in the periplasm. The first and last strands of and the periplasmic turns T1-T6. In detergent micelles, BamA is ob-BamA (β1and β16) creates a lateral gate, which acts as the functional served mostly in the inward-open conformation. The native mem-hotspot for protein folding. Structural studies have revealed three brane considerably modulated the conformational space. BamA major conformations of the BamA barrel namely inward open (IO), exists as a monomer and excurses over a broad range of conforma-lateral open (LO), and lateral open substrate-bound states [1,2]. In tions including the inward open, lateral open, and lateral open sub-the IO state, the β16 strand exists in a fully zipped conformation, strate-bound states. Our results validate the three major conforma- INTRODUCTION whereas in the LO, it is in a kinked state. The lateral opening further tions of BamA in the cellular membrane. In summary, BamA alone SPONSORS increases when the substrate is bound. Only BamA and BamD are can transverse through multiple conformations without any of the PROGRAM essential and the role of other lipoproteins or how the conforma- lipoproteins, which might be the crucial aspect underlying its diver- PROGRAM — BY DAY tional dynamics of the complex is regulated remains unknown. The gent functions in vivo. TOPICS outer membrane is an asymmetric bilayer consisting of phospho- PRIZE LECTURES lipids and lipopolysaccharides. It has been impossible to replicate PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] Y. Gu, H. Li, H. Don, et al, Nature. 2016, 531, 64–69. [2] D. Tomasek, S. Rawson, J.Lee, et al, Nature. 2020, 120, 356-365. [3] B. Joseph, et al, Nat. Protoc. 2019, 14(8):2344-2369. PO126 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 279 ANNEMARIE KEHL1, RESOLUTION OF CHEMICAL SHIFT ANISOTROPY IN 19F MARKUS HILLER1, ENDOR SPECTROSCOPY AT 263 GHZ/9.4 TESLA IGOR TKACH1, SEBASTIAN DECHERT2, Fluorine spin labels have found many applications in magnetic resonance due to their favorable properties. Partic-MARINA BENNATI1,2, ularly, the nuclear spin of 1/2, 100% natural abundance, a high gyromagnetic ratio and a large range of chemical ANDREAS MEYER1 shifts make them valuable in NMR and high field ENDOR.[1,2] Our quasi-optical EPR spectrometer enables ENDOR 1Max Planck Institute for Biophysical measurements at 263 GHz with field strengths comparable to NMR (9.4 T), which increase spectral resolution and Chemistry, Research Group EPR orientation selectivity.[3] Spectroscopy, Göttingen, Germany 2University of Göttingen, Department In this contribution 19F ENDOR spectra at 263 GHz and 94 GHz are compared for five nitroxide radical model systems of Chemistry, Göttingen, Germany consisting of a phenyl group substituted with fluorine or a CF -group. The spectra at 263 GHz display asymmetry, 3 which is not resolved in the spectra at 94 GHz. It indicates that the chemical shift anisotropy (CSA) is resolved, which so far has been neglected in EPR and ENDOR spectroscopy. Analysis of the spectra both at 263 GHz and 94 GHz is presented. For the simulations, the orientation of the chemical shift tensor towards the g-frame is described by Euler angles and can provide further structural information on the sample. The CSA can be resolved also for difluorinated samples as long as the hyperfine coupling tensors are resolved. For the CF substituted system the resolution of chemical shift is possible in case free rotation of the group 3 is hindered, however, analysis relys on DFT calculations due to the many interdependent parameters. In comparison to 94 GHz, 19F ENDOR at 263 GHz can display high resolution spectra and can improve spectral inter- INTRODUCTION pretation by providing information on the chemical shift tensor. SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] K. E. Arntson, W. C. K. Pomerantz,. J. Med. Chem. , 2016, 59, 5158-5171. [2] A. Meyer, S. Dechert, S. Dey, C. Höbartner, M. Bennati, Angew. Chem. ADVERTISERS Int. Ed. , 2020, 59, 373-379. [3] I. Tkach, I. Bejenke, F. Hecker, A. Kehl, M. Kasanmascheff, I. Gromov, I. Prisecaru, P. Höfer, M. Hiller, M. Bennati, JMR, 2019, AUTHOR INDEX 303, 17 – 27. PO127 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 280 SHINY MAITY*1, TOWARD TIME-RESOLVED GD-GD DIPOLAR EPR FOR TRIGGERED BRAD PRICE*1, C. BLAKE FUNCTIONAL DYNAMICS IN PROTEINS WILSON1, CHUNG-TA HAN1, XIAOLING WANG1, Protein based molecular machines are central to every biological pro- changes in residue local environment in real time [3]. Taking advantage ARNAB MUKHERJEE1, cess and enable living cells to perform basic functions. Our under- of the well-known effects of dipolar coupling on EPR linewidth, TiGGER MAXWELL WILSON1, standing of these machines is primarily based on their 3D structure -- a will collect linewidth information of the protein sample every 10µs that JANET LOVETT2, MATHIEU static picture. In order to understand their operation as machines it is can be used to determine inter-spin (and therefore, residue-residue) STARCK3, DAVID PARKER3, necessary to investigate the evolution of their conformational changes distance as a function of time. Further, due to the exceptionally narrow SONGI HAN1, MARK in time, triggered by an external stimulus. The combination of site-di- line of Gd(III) labels at high field (~8.57 T) [4], a low rapid-scan sweep SHERWIN1 rected spin labeling (SDSL) and electron paramagnetic resonance (EPR) rate will avoid rapid passage effects while still collecting the full line- 1UC Santa Barbara, Department of Chemistry is a well-established biophysical tool that has been used to study light shape in a matter of microseconds [5]. and Biochemistry, Santa Barbara, CA, USA 2 activated conformational changes in proteins in real time at room tem- University of St. Andrews, School of Physics We chose to focus on a light-sensitive protein called AsLOV2 which and Astronomy, St. Andrews, Scotland, UK perature, which can also be correlated with chromophore-sensitive 3 has gained great popularity in the bioengineering field as mechan- University of Durham, Department of photocycle kinetics [1]. We are developing time-resolved (µs-s times- Chemistry, Durham, UK ical actuators, because they offer control of cellular activities with cales) Gd-Gd dipolar-broadening EPR (TiGGER) for tracking protein spatiotemporal specificity. The widespread application of these functional dynamics in a close-to-native environment. proteins has motivated efforts to engineer improved variants [6,7], We will present our work toward TiGGER, which includes progress in making it necessary to understand the long-range non-equilibrium high efficiency double-spin labeling of a light-sensitive protein, as well structural changes [8] that accompany AsLOV2 on light activation. as hardware design and testing. TiGGER makes use of the ability of 240 We will apply TiGGER to elucidate important time-dependent struc- INTRODUCTION GHz Gd(III) cwEPR to extract inter-spin distances exceeding 2 nm at tural information on AsLOV2, as well as other optogenetic mecha- SPONSORS room temperature [2] and will utilize rapid-scan transient EPR to track nosensitive proteins in the future. PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE References: [1] R. Mollaaghababa, et al., Biochemistry 2000, 39(5), 1120-1127. [2] J.A. Clayton, et al., Physical Chemistry Chemical Physics 2017, 19(7), 5127-5136. [3] C.B. Wilson, Adventures in INVITED AND PROMOTED LECTURES high-field electron paramagnetic resonance, UC Santa Barbara, 2019. [4] J.W. Stoner, et al., Journal of Magnetic Resonance 2004, 170(1), 127-135. [5] M. Tseitlin, G. A. Rinard, R. W. Quine, S. S. POSTERS Eaton, & G. R. Eaton, Journal of magnetic resonance 2011, 208(2), 279-283. [6] D. Strickland, et al., Nat. Methods 2010, 7, 623–626. [7] A. Mukherjee, et al., Journal of Biological Engineering 2012, ADVERTISERS 6, 20. [8] S. Harper, L. Neil, and K. Gardner, Science 2003, 301, 1541–1544. AUTHOR INDEX Acknowledgements: We gratefully acknowledge financial support from NSF MCB-2025860 and MRI-19-601107. PO128 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 281 HANNAH RUSSELL1, DEER AND RIDME MEASUREMENTS OF RACHEL STEWART1, THE NITROXIDE-SPIN LABELLED CHRISTOPHER PRIOR2, VASILY S. OGANESYAN2, COPPER-BOUND AMINE OXIDASE HOMODIMER FROM THEMBANINKOSI ARTHROBACTER GLOBIFORMIS G. GAULE3, JANET E. LOVETT1 In the study of biological systems, pulse dipolar spectroscopy (PDS) methods are known to be invaluable techniques 1SUPA School of Physics and capable of measuring dipolar-dipole interactions between paramagnetic centres which, in turn, aid the elucidation Astronomy and BSRC, University of of nanometre-scale distances. In this work, Double Electron Electron Resonance (DEER) and Relaxation Induced St Andrews, St Andrews, UK 2School of Chemistry, University of Dipolar Modulation Enhancement (RIDME) experiments are carried out on the copper amine oxidase from Arthro- East Anglia, Norwich, UK bacter globiformis (AGAO), which contains multiple measurable nanometre-scale distances between nitroxide spin 3School of Molecular and Cellular labels and bound Cu(II) paramagnetic centres. To aid in the identification of the distances measured within the Biology, Astbury Centre for Structural Molecular Biology, AGAO homodimer, protein modelling techniques are employed. Using these models for comparison to experimen- University of Leeds, Leeds, UK tal data, we determine that the distances measured within the homodimer indicate that the active Cu(II) binding sites are unoccupied, and instead, the Cu(II) is binding to surface sites. We show that two distinct distances can be measured, with the longer of the two providing a measurement that exceeds 5 nm, through optimisation of experimental parameters and careful data analysis, and using these results, we also discuss that the use of RIDME experiments may hold advantages over DEER for measurement of these long distances [1]. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX References: [1] H. Russell, R. Stewart, C. Prior, V.S. Organesyan, T.G. Gaule, J.E. Lovett, Appl. Magn. Reson. 2021, 1-21. PO129 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 282 PAUL TRENKLER, INVESTIGATIONS ON THE RELAXATION BEHAVIOUR OF ANDREI KUZHELEV, NITROXIDES AT HIGH FIELDS AND PREDICTIONS OF THE THOMAS F. PRISNER Institute of Physical and Theoretical RESULTING RIDME BEHAVIOUR Chemistry and Center for Biomolecular Resonance, Goethe University Frankfurt, Pulsed dipolar spectroscopy (PDS) includes multiple methods in with bis-nitroxides at high magnetic fields could be. Therefore we Frankfurt am Main, Germany electron paramagnetic resonance (EPR) that allow to extract dis- investigated the relaxation behavior of nitroxides in dependence tance information from the dipolar interaction between paramag- of different parameters, such as magnetic field strength B , sample 0 netic spin labels in a 2-8 nm range. By far the most popular PDS temperature T, radical concentration c and mixing time T in a mix technique is pulsed electron-electron double resonance (PELDOR RIDME experiment. also called DEER). But particularly at higher fields, where the g-an- Very long phase-memory times T between 20-30 µs where mea- isotropy causes spectra to be extremely broad, the efficiency of PEL- m sured at 263 GHz for two different nitroxides at a concentra- DOR is severely limited by the small available excitation bandwidth tion of 0.5 mmol/L in a deuterated solvent matrix. Even higher of high frequency pulses. phase-memory times can be expected by further reducing the A PDS technique which is less affected by the small microwave pulse concentration, since an increase of 10 µs was observed with halve excitation bandwidth is Relaxation-induced dipolar modulation of the concentration. While these values predict a high potential of enhancement (RIDME). In contrast to PELDOR, RIDME does not use the RIDME method for nitroxide spin-labels, we also realized that a a microwave pulse for the inversion of the dipolar coupled spin, but possible limitation for the RIDME efficiency at high fields might be relies on relaxation processes and is therefore independent of the an enlarged spectral diffusion contribution to the relaxation times. INTRODUCTION available excitation bandwidth. This should make RIDME particu- Inspired by the paper from Ritsch et al. [2] we propose a method SPONSORS larly suited for high field measurements. for estimating optimal temperatures and mixing times in a RIDME PROGRAM In the past we have shown that RIDME with high spin Mn2+ centers experiment for bis-nitroxides. We also consider the high spectral PROGRAM — BY DAY gives both higher modulation depths and higher signal-to-noise ra- anisotropy of T and an effective memory time T as a function of 1 m TOPICS tio compared to PELDOR at 260 GHz microwave frequency.[1] In the the mixing time T for the calculation of the RIDME experiment mix PRIZE LECTURES study presented here, we were interested in how efficient RIDME efficiency. PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS References: [1] D. Akhmetzyanov, H.Y.V. Ching, V. Denysenkov, P. Demay-Drouhard, H.C. Bertrand, L.C. Tabares, C. Policar, T.F. Prisner and S. Un, Phys. Chem. Chem. Phys. 2016, 18, 30857- AUTHOR INDEX 30866. [2] I. Ritsch, H. Hintz, G. Jeschke, A. Godt, M. Yulikov, Phys. Chem. Chem. Phys. 2019, 21, 9810-9830. PO130 POSTERS / EPR IN BIOMOLECULAR AND MATERIAL SCIENCE 283 CEZARA ZAGREAN-TUZA1, THROUGH THE LOOKING-GLASS: GRIGORE DAMIAN2, A GLIMPSE AT HOW SMALL LIGANDS MODULATE THE MAGNETIC AUGUSTIN C. MOT1, RADU SILAGHI-DUMITRESCU1 PROPERTIES OF LOW-SPIN HEMOGLOBIN ADDUCTS 1Department of Chemistry, Babes-Bolyai University, Cluj-Napoca, Romania In the beginning there was the protein, and the protein was hemo- broadening is observed for low-spin complexes [2], where normally 2Department of Physics, Babes-Bolyai globin, but not the ferric type. Even though it is probably the best spin-lattice relaxation has no effects whatsoever - a phenomenon University, Cluj-Napoca, Romania known protein, hemoglobin still continues to unfold new roles and known as g strain: the linewidth broadens as a result of random dis-properties, ranging from physiological implications and moonlight- tribution of protein conformations. ing to nanotechnological applications. In the early days of biochem- The present work explores the behaviour of various small strong- istry, the Fe3+ form of hemoglobin was deemed a useless byproduct field ligands in their binding to ferric hemoglobin from the perspec-since (by contrast to the Fe2+ form) it has no ability to bind molecular tive of the broadening effect in CW-EPR. From this ligand series, oxygen. It was later understood that a large number of heme-con- some structurally-related small moieties stand out as imposing a taining enzymes in fact rely primarily on their ferric forms to per- longer relaxation time, thus rendering a violation from the g strain form catalysis (e.g., catalase, ascorbate peroxidase, manganese per- rule. Among these, one small molecule, used in the past to treat oxidase, cytochrome P450NOR). Ferric heme proteins additionnally alcohol poisoning, goes beyond all expectations and gives rise to have an increased ability (compared to their ferrous counterparts) a very rare methemoglobin EPR signal, where the expected broad-to bind probe ligands - from very small ones such as cyanide, to the ening is almost absent. The observed paramagnetic behaviours are larger molecules such as anthracene as seen in cytochromes P450. explored by use of molecular modeling approaches in trying to un- INTRODUCTION Starting with the pioneering work of Peisach and Blumberg, derstand which factors are responsible for such a g-strain-ignorant SPONSORS through Taylorś formulae for calculation of ligand field parame- EPR manner. If you are interested to know why some low-spin heme PROGRAM ters [1], interest has been paid in understanding how various small adducts can become so impolite as to refuse to abide the rules, stay PROGRAM — BY DAY ligands affect the magnetic properties of hemoglobin in EPR spec- tuned and check out the poster. TOPICS troscopy. Even at low temperatures such as 70 K, an unusually large PRIZE LECTURES PLENARY LECTURES TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] C. P. S. Taylor, Biochim. Biophys. Acta. 1977, 491.1, 137-148. [2] A. I. Ioanitescu, et al., Biophys J. , 2005, 89.4, 2628-2639. ADVERTISERS Acknowledgements: This work was partly supported by a grant of the Romanian Ministry of Research, Innovation and Digitization, CNCS/CCCDI—UEFISCDI, project number TE-2019-1396, AUTHOR INDEX within PNCDI III. PO131 POSTERS / FRONTIERS IN MAGNETIC RESONANCE 284 ALEXANDER BARNES1, ELECTRON DECOUPLING ON ENDOFULLERENES AND EDWARD SALIBA2, MAS ELECTRON SATURATION RECOVERY ERIKA L. SESTI3, PATRICK T. JUDGE4, Presented here is a demonstration of control over electron spins resolution of DNP-enhanced nuclear spins.[3] The longer electron NICHOLAS ALANIVA1 in two different systems: 40 mM Trityl and nitrogen-endofuller- spin relaxation at lower temperatures (90 K, and especially below 1ETH-Zürich, Department of ene (N@C60). The electron saturation recovery of the unpaired 5 K) improves our ability to control electron spins with frequen-Chem. and Appl. Biosci., Zürich, electron spins in 40 mM Trityl is observed through the enhanced cy-modulated microwaves. Switzerland 2Massachusetts Institute of nuclear magnetic resonance (NMR) signal of proline, using a rap- Here, electron decoupling provides electron saturation for “direct” Technology, Cambridge, MA, USA id-acquisition scheme involving electron decoupling and direct 3Pfizer, Inc., Chesterfield, MO, USA DNP experiments, where the polarization period varies from milli- 4 dynamic nuclear polarization (DNP), while magic-angle spinning Washington University in St. Louis, seconds to tens-of-milliseconds, after which the longitudinal recov- St. Louis, MO, USA (MAS) at temeratures below 5 K. Additionally, electron decou- ery of the electron spin (40 ms) is observed through the enhanced pling is extended to a dilute electron-spin system, polycrystalline nuclear spin signal.[4] To successfully encode this electron spin in-N@C60: C60-fullerene (160 parts-per-million (ppm) endo-species), formation in the nuclear signal, the longitudinal relaxation of the to improve signal intensity. 40 mM Trityl electron spins must be longer than the DNP transfer DNP is a process whereby the relatively high polarization of elec- period, thus MAS is performed at 5 K using liquid helium for cool- tron spins is transferred to nuclear spin; here, the transfer is accom- ing. A rapid-acquisition scheme optimizes the signal-per-unit-time plished with microwaves from a custom frequency-agile gyrotron. of the spectrometer, allowing for the necessary signal acquisition at [1-2] MAS NMR is a technique used to improve the senstivity and these short polarization periods. resolution of solid state NMR, making possible accurate determi- INTRODUCTION In contrast to 40 mM trityl, which features some radical-clusters, nation of structrue and dynamics in a wide range of low-mobility SPONSORS 160 ppm N@C60 provides an environment with essentially no elec- systems, from inorganic surfaces to in-cell proteins. DNP drasti- PROGRAM tron-electron interaction. In this electron-spin-dilute system we cally decreases the time required for these information-rich ex- PROGRAM — BY DAY successfully perform electron decoupling, observing a 12% increase periments, but unfortunately the electron-nuclear dipolar contact TOPICS in signal intensity at 7 s DNP period and a 5% increase in signal in- utilized for signal enhancement also broadens the NMR signal, re- PRIZE LECTURES tensity at 30 s DNP period (for the isotropic peak).[8] sulting in spectra with poor resolution. Electron decoupling attenu- PLENARY LECTURES ates this electron-nuclear interaction and recovers the intensity and TUTORIAL LECTURE INVITED AND PROMOTED LECTURES POSTERS References: [1] A. Barnes, et al., Appl. Mag. Res. 2008, 34. [2] N. Alaniva, C. Gao, et al., JMR. 2019, 308. [3] E. Sesti, et al., JMR. 2018, 295. [4] N. Alaniva, E. Saliba, et al., Angew. Chemie. 2019, ADVERTISERS 22. [5] N. Alaniva, et al., in progress AUTHOR INDEX Acknowledgements: Björn Corzilius, Snorri Th. Sigurdsson, Chukun Gao. 285 THE EUROMAR 2021 IS MADE POSSIBLE THROUGH THE GENEROUS SUPPORT OF OUR SPONSORS Take Your Research to the Next Level Bruker GHz-class NMR spectrometers allow for advanced research on structural-functional biology of proteins and protein complexes including intrinsically disordered proteins (IDPs). The Bruker line of 1.1 and 1.2 GHz NMR instruments provide unsurpassed stability and spectral resolution allowing researchers to see further than ever before. INTRODUCTION SPONSORS PROGRAM PROGRAM — BY DAY TOPICS PRIZE LECTURES PLENARY LECTURES To learn more about our products visit our web- TUTORIAL LECTURE site or e-mail us at: INVITED AND PROMOTED LECTURES POSTERS ADVERTISERS AUTHOR INDEX NMR Innovation with Integrity MORE THAN 60 YEARS MANUFACTURING EXCELLENCE IN NMR Learn more about Innovations in JEOL NMR Technology Workshop 1 : July 5th: Meet the JEOL Team User Meeting : July 6th: “Linking structure and magnetic properties in minerals using solid state NMR and other techniques” – by Ulla Gro Nielsen, (University of Southern Denmark). OUR KNOWLEDGE FOR YOUR HEALTH. “NMR crystallography of molecular solids” – by Martin Drac í nsky , (Czech Academy of Sciences). Top quality products are our commitment and Workshop 2 : July 7th: “Pulse Shaping in Delta” – people’s health is our motivation. by Evgeny Tishchenko, (JEOL USA). Krka is one of the world’s leading generic pharmaceutical companies. 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Contact us today for a demonstration The life science The life science business of Merck business of Merck operates as operates as nmr.oxinst.com/x-pulse MilliporeSigma in MilliporeSigma in the U.S. and Canada. the U.S. and Canada. The life science business of Merck operates as MilliporeSigma in the U.S. and Canada. CJ1117 X-Pulse Broadband Ad 146 X 210.indd 1 15/06/2021 12:48 The right chromatography tools for the right tasks - and the skilst ou set hem. T H A T ’ S W H O W E A R E . BIA Separations is the leading developer of monolith technology and the exclusive producer of CIM® (convective interaction media) chromatographic columns for the production, purification, and analysis of large biomolecules. www.biaseparations.com Browser-Based NMR Data Processing and Analysis Process 1D and 2D NMR data in your browser – no individual software downloads required! Spectrus JS: Coming this summer! ® 50% Academic Discount until August 31, 2021 Learn More: www.acdlabs.com/SpectrusJSPromo Head Office: Worldwide Offices: https://www.youtube.com/channel/UCE1IlXjIIorZKchjvd3Vtng Short educational videos about Magnetic Resonance curated by the Groupement AMPERE. Interested in contributing with your own video? Contact: videos@ampere-society.org AMPERE Video Committee: Matthias Ernst, Alvar Gossert, Sebastian Hil er, Lauriane Lecoq, Oscar Mil et, Yi-Qiao Song 295 AUTHOR INDEX AUTHOR INDEX 296 PRIZE, A. A. Mulder, Frans IN040 Coudevylle, Nicolas PT039 PLENARY, Aebischer, Kathrin PT029 Di Gregorio, Enza PT035 TUTORIAL, Agustin Romero, Javier PT024 Dorai, Kavita IN051 INVITED AND Alderson, T. Reid PR002 Dračínský, Martin PT060 PROMOTED Ardenkjær-Larsen, Jan Henrik PR003 Dumez, Jean-Nicolas IN011 Bagryanskaya, Elena PT041 Elena-Herrmann, Benedicte IN050 Baldus, Marc IN005 Emsley, Lyndon PL015 Batta, Gyula PT031 Falourd, Xavier PT056 Beira, Maria PT013 Farjon, Jonathan PT030 Bernado, Pau IN030 Ferrage, Fabien PL005 Bielskute, Stase PT032 Franssen, Wouter PT034 Blümich, Bernhard IN022 Frydman, Lucio PR004 Bode, Bela IN032 Frydman, Lucio PL007 Bordignon, Enrica PL010 Garbacz, Piotr PT001 Brauser, Matthias PT048 Gauger, Maximilian PT043 Britton, Melanie IN027 Geninatti Crich, Simonetta PT014 INTRODUCTION SPONSORS Brutscher, Bernhard IN020 Georgoulia, Panagiota PT033 PROGRAM Budker, Dmitry IN023 Gil, Ana PL014 PROGRAM — BY DAY Butts, Craig PL004 Glaubitz, Clemens IN048 TOPICS Camacho Zarco, Aldo PT047 Gosar, Žiga PT006 PRIZE LECTURES PLENARY LECTURES Carlomagno, Teresa PL009 Halse, Meghan IN017 TUTORIAL LECTURE Claridge, Tim IN042 Hiller, Markus PT042 INVITED AND PROMOTED LECTURES Cordova, Manuel PT061 Hiller, Sebastian PT044 POSTERS Coronado, Eugenio PL012 Himmler, Aaron PT023 ADVERTISERS AUTHOR INDEX Corzilius, Björn IN016 Horvatić, Mladen IN003 AUTHOR INDEX 297 Imai, Takashi PT004 Meichsner, Shari PT007 J. Kubicki, Dominik IN015 Mendels, Philippe PL002 J. Pielak, Gary PL003 Mentink-Vigier, Frederic PT020 J. Rossini, Aaron IN019 Menzildjian, Georges PT021 Jannin, Sami PT019 Mileo, Elisabetta IN006 Jelezko, Fedor IN043 Mishra, Rituraj PT010 Jimenez-Barbero, Jesus IN037 Mompean, Miguel PT054 Juramy, Marie PT011 Mott, Helen IN002 Kazimierczuk, Krzysztof PT027 Nassrin Kriebel, Clara PT055 Kimmich, Rainer TU001 Nawrocka, Ewa PT058 Korenchan, David PT052 Neugebauer, Petr IN031 Krajnc, Andraž IN053 Nilsson, Mathias IN009 Kruk, Danuta IN012 Nouri, Sirine PT003 Kurzbach, Dennis IN018 Novak, Predrag IN041 Lafon, Olivier PT015 Oosterkamp, Tjerk IN045 Leskes, Michal IN014 Ovcherenko, Sergey PT008 INTRODUCTION SPONSORS Li, Qiang PT002 P.M. Kentgens, Arno PL006 PROGRAM Loquet, Antoine PR001 Perczel, Andras IN033 PROGRAM — BY DAY Lu, Jiaqi PT009 Pérez Trujillo, Míriam PT059 TOPICS Luchinat, Enrico IN007 Petzold, Katja IN039 PRIZE LECTURES PLENARY LECTURES Lurie, David IN013 Pierattelli, Roberta PL001 TUTORIAL LECTURE M. Thiele, Christina IN010 Piersanti, Elena PT050 INVITED AND PROMOTED LECTURES Madl, Tobias IN029 Pintacuda, Guido PL013 POSTERS Mao, Jiafei PT028 Pons, Miquel IN025 ADVERTISERS AUTHOR INDEX Mavromoustakos, Thomas IN034 Pylaeva, Svetlana PT018 AUTHOR INDEX 298 Raftery, Daniel IN049 Weingarth, Markus IN047 Randazzo, Antonio IN035 Wisser, Dorothea PT017 Ribay, Victor PT049 Wolf, Tamar PT016 Riek, Roland IN001 Wrachtrup, Joerg IN044 Ringkjøbing Jensen, Malene IN026 Wu, Bing PT062 Rovó, Petra IN038 Zangger, Klaus PT025 Schiemann, Olav IN008 Zhukov, Ivan PT026 Schmidt, Thomas PT040 Židek, Lukaš IN024 Schmidt-Rohr, Klaus IN052 Zorko, Andrej IN004 Schwalbe, Harald IN036 Zvyagin, Sergei PT005 Sessoli, Roberta IN046 Shemesh, Noam PL008 Simões de Almeida, Bruno PT053 Smith, Albert PT045 Štěpánek, Petr PT051 Stoll, Raphael PT038 INTRODUCTION SPONSORS SubbaRao Redrouthu, Venkata PT022 PROGRAM Sykora, Jan PT057 PROGRAM — BY DAY Tayler, Michael PT012 TOPICS Telkki, Ville-Veikko IN021 PRIZE LECTURES PLENARY LECTURES Toerner, Ricarda PT037 TUTORIAL LECTURE Tomlinson, Jennifer PT046 INVITED AND PROMOTED LECTURES Tušar, Kaja PT036 POSTERS Van Doorslaer, Sabine PL011 ADVERTISERS AUTHOR INDEX Węglarz, Władysław IN028 AUTHOR INDEX 299 POSTER Aleksič, Simon PO002 Bispo, Daniela S. C. PO069 Bracaglia, Lorenzo PO003 Bizkarguenaga, Maider PO059 Gómez, M. Victoria PO084 Bretschneider, Matthias PO104 Gupta, Agrim PO007 Bruzzone, Chiara PO070 Klukowski, Piotr PO009 Brzezicki, Artur PO048 Kocman, Vojč PO010 Burakova, Ekaterina PO029 Kovačič, Matic PO011 Burkhard , Endeward PO124 Lenarčič Živković, Martina PO012 Callon, Morgane PO030 Levstek, Tevž PO013 Carneiro, Tatiana J. PO071 P. Kalverda, Arnout PO008 Cevec, Mirko PO004 A. Słowiński, Mateusz PO113 Chávez, Matías PO105 Adams, Alina PO117 Chen, Xiaoxun PO122 Adler , Agnes PO001 Chontzopoulou, Eleni PO060 Alaniva, Nicholas PO131 Denysenkov, Vasyl PO082 Andriy, Marko PO087 Derks, Maik PO031 Araújo, Rita PO068 Di Cesare, Francesca PO072 INTRODUCTION SPONSORS Argyropoulos, Dimitris PO046 Dispinar Gezer, Tugba PO118 PROGRAM Asanbaeva, Nargiz PO121 Dolenc, Katja PO061 PROGRAM — BY DAY Ausmees, Kerti PO079 Duarte, Daniela PO073 TOPICS Azagra, Marc PO102 Duchowny, Anton PO119 PRIZE LECTURES PLENARY LECTURES Badoni, Saumya PO080 Elkhanoufi, Sabrina PO123 TUTORIAL LECTURE Bayzou, Racha PO103 Elliott, Stuart J. PO083 INVITED AND PROMOTED LECTURES Bazzoni, Margherita PO047 Feguš, Nastja PO005 POSTERS Bernardi, Marie PO091 Ferrante, Gianni PO092 ADVERTISERS AUTHOR INDEX Biedenbänder, Thomas PO081 Fridolf, Simon PO032 AUTHOR INDEX 300 Gajarsky, Martin PO006 Mihelač, Mateja PO054 Gansmüller, Axel PO099 Mikulandra, Ivana PO063 Gołowicz, Dariusz PO106 Mishra, Aditya PO097 Gopinath, Aathira PO125 Mohan, Megha PO109 Gradišek, Anton PO093 Morales-Melgares, Anna PO096 Habinovec, Iva PO049 Negi, Deepraj PO016 Horník, Štěpán PO074 Novotný, Aleš PO017 Kachariya, Nitin PO037 Obkircher, Markus PO055 Kehl, Annemarie PO126 Orfanidis, Savvas PO100 Kervern, Gwendal PO107 Osifová, Zuzana PO056 Ketter, Sophie PO041 Pavc, Daša PO018 Kolar, Maja PO062 Pečnik, Klemen PO075 Krafčik, Daniel PO042 Pérez, Yolanda PO064 Kristinaityte, Kristina PO050 Peterková, Kateřina PO019 Kumar, Anant PO094 Podbevšek, Peter PO020 Kumar Reddy Sannapureddi, Rajesh PO039 Pokharna, Aditya PO021 INTRODUCTION SPONSORS Kumar Sreemantula, Arun PO033 Pontoriero, Letizia PO022 PROGRAM Levien, Marcel PO086 Pustovalova, Yulia PO065 PROGRAM — BY DAY Liu, Kristina S. PO108 Rangadurai, Atul PO038 TOPICS Maity, Shiny PO127 Rathner, Adriana PO023 PRIZE LECTURES PLENARY LECTURES Makuc, Damjan PO051 Reimets, Nele PO088 TUTORIAL LECTURE Makulski, Włodzimierz PO052 Renou, Sophie PO110 INVITED AND PROMOTED LECTURES Martek, Bruno Aleksander PO053 Ridvan, Ince PO111 POSTERS Marušič, Maja PO014 Russell, Hannah PO128 ADVERTISERS AUTHOR INDEX Medved, Nataša PO015 Ryneš, Jan PO043 AUTHOR INDEX 301 S. Chakrabarti, Kalyan PO036 Wang, Zhuoran PO090 Šadl, Alen PO025 Weiss, Charlotte PO034 Safeer, Adil PO044 Wurl, Anika PO035 Sathyamoothy, Bharathwaj PO024 Yves, Gossuin PO120 Scheler, Ulrich PO101 Zagrean-Tuza, Cezara PO130 Schiavina, Marco PO066 Zoupanou, Nikoletta PO067 Sebastião, Pedro J. PO095 Šedivý, Matúš PO116 Shchukina, Alexandra PO112 Simova, Svetlana PO076 Šket, Primož PO026 Škrjanc, Monika PO077 Solodovnyk, Artur PO114 Srivastava, Deepansh PO115 Stern, Quentin PO089 Štih, Vesna PO027 INTRODUCTION SPONSORS Svenningsson, Leo PO057 PROGRAM Taylor, Michael J. PO058 PROGRAM — BY DAY Trajkovski, Marko PO028 TOPICS Trenkler, Paul PO129 PRIZE LECTURES PLENARY LECTURES Vignoli, Alessia PO078 TUTORIAL LECTURE Vinod Kumar, Sanjay PO085 INVITED AND PROMOTED LECTURES Víšková, Pavlína PO045 POSTERS Volavšek, Janez PO098 ADVERTISERS AUTHOR INDEX Wang, Ying PO040 PUBLISHER: EN-FIST CENTRE OF EXCELLENCE LJUBLJANA, SLOVENIA, JULY, 2021 EDITORS: MARTINA LENARČIČ-ŽIVKOVIĆ MAJA MARUŠIČ JANEZ PLAVEC PETRA JAKSETIČ EXECUTIVE CONFERENCE PRODUCTION: EVENT SERVICE GROUP / MBZ D.O.O. GRAPHIC DESIGN & LAYOUT: HERCOGMARTINI NUMBER OF COPIES: 50 CIP - Kataložni zapis o publikaciji Univerzitetna knjižnica Maribor 577.1/.2:601.2:543.429.23(082) EUROMAR. European Conference on Magnetic Resonance (2021 ; Ljubljana) EUROMAR : European Conference on Magnetic Resonance, 5th - 8th July, 2021, Ljubljana : book of abstracts / [editors Martina Lenarčič-Živkovič ... [et al.]. - Ljubljana : En-Fist Centre of Excellence, 2021 ISBN 978-961-95442-0-4 COBISS.SI-ID 68668163