title: Purification and characterization of a protease from Xenopus embryos.
abstract: A proteolytic enzyme was purified from Xenopus embryos. The purification procedure consisted of fractionation of an extract of embryos with acetone, gel filtration of Sephadex G-75 and chromatography on carboxymethyl-cellulose and hydroxylapatite. The preparation of enzyme appeared to be homogeneous as judged by electrophoresis in polyacrylamide gels. This protease had a molecular mass of 43-44 kDa and was composed of two subunits with molecular masses of 30 kDa and 13 kDa. The optimal pH of the reaction catalysed by the protease was approximately 4.0. This proteolytic activity was inhibited by antipain, leupeptin and iodoacetic acid; it was not affected by phenylmethylsulfonyl fluoride and pepstatin; and it was enhanced by dithiothreitol. In the presence of RNA, the optimal pH was shifted from pH 4.0 to pH 4.5. The protease was activated by addition of total RNA from Xenopus embryos, by poly(rU) or poly(rG). In contrast, after addition of tRNA or poly(rC), no activation of the protease was observed.
authors: Laboratory of Research for Biosynthesis and Metabolism, Keio University, School of Medicine, Tokyo, Japan.
authors: S
authors: S
authors: Miyata
authors: Y
authors: Y
authors: Yoshida
authors: H K
authors: HK
authors: Kihara
mesh_terms: D000818:Animals; D000976:Antipain / Q000494:pharmacology; D002845:Chromatography; D004625:Embryo, Nonmammalian / Q000201:enzymology; D004789:Enzyme Activation; D007461:Iodoacetates / Q000494:pharmacology; D019807:Iodoacetic Acid; D007976:Leupeptins / Q000494:pharmacology; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000096:biosynthesis / Q000302:isolation & purification*; D011131:Polyribonucleotides / Q000494:pharmacology; D012313:RNA / Q000494:pharmacology; D013379:Substrate Specificity; D014981:Xenopus
chemical_list: D007461:Iodoacetates; D007976:Leupeptins; D011131:Polyribonucleotides; D000976:Antipain; D012313:RNA; D010447:Peptide Hydrolases; C032854:leupeptin; D019807:Iodoacetic Acid