15 KB121 KB14 KB199820252009Kos, JankoPečar Fonović, Uršaštevilka:4letnik:56str. 985-988ISSN:1318-0207COBISSID:2730353URN:URN:NBN:SI:doc-28GNWRQ0enSlovensko kemijsko društvoActa chimica slovenicabiokemijacisteinske proteazečiščenjeizolacijakatepsinEfficient removal of cathepsin L from active cathepsin X using immunoprecipitation technique|Cathepsin X is a cysteine protease which is involved in various important physiological and pathological processes. For the purpose of biochemical and structural studies, pure and active cathepsin X is required without contamination with other related proteases. Recombinant cathepsin X was obtained by expression in methylotropic yeast Pichia pastoris. During purification, cathepsin X has to be activated with cysteine protease cathepsinL, however, separation methods, used so far, can not completely remove cathepsin L from the activated cathepsin X. Here we describe a new purification procedure which provides active recombinant cathepsin X without the presence of residual cathepsin LKatepsin X je cisteinska proteaza, ki je udele`ena v razli~nih pomembnih fiziolo{kih in patolo{kih procesih. Za potrebe biokemijskih in strukturnih raziskav je potreben ~ist in aktiven katepsin X brez vsebnosti sorodnih proteaz. Rekombinantni katepsin X smo izrazili v metilotropni kvasovki Pichia pastoris. V postopku izolacije je potrebno katepsin X aktivirati s cisteinsko proteazo katepsinom L, vendar z do sedaj objavljenimi metodami ni bilo mogo~e popolnoma lo~iti katepsina L in aktivnega katepsina X. V ~lanku opisujemo novometodo izolacije, s katero dobimo aktiven katepsin X brez prisotnosti katepsina LTEXTznanstveno časopisjejournalsSlovenian National E-content AggregatorNational and University Library of SloveniaSlovensko kemijsko društvo