家蚕卵半胱氨酸蛋白酶的分离、纯化及性质研究

Isolation, purification, and properties of cysteine proteinase from Bombyx mori L. eggs.

作者信息

Yarygin D V, Klunova S M, Filippovich Yu B

机构信息

Department of Organic and Biological Chemistry, Moscow State Pedagogical University, Moscow, 129278, Russia.

出版信息

Biochemistry (Mosc). 2003 Jan;68(1):63-7. doi: 10.1023/a:1022145518691.

DOI:10.1023/a:1022145518691

PMID:12693978

Abstract

Silkworm moth (bombyx) egg cysteine proteinase with maximal activity at pH 3.0 was purified by chromatography and isoelectrofocusing. On SDS-electrophoresis in polyacrylamide gel the purified enzyme showed a single band of molecular mass 50 kD. Isoelectrofocusing revealed that the bombyx egg cysteine proteinase exists in two forms with pI values of 5.95 and 6.43, respectively. The enzyme activity was sensitive to inhibition by iodoacetamide and p-chloromercuribenzoate but resistant to EDTA, pepstatin, and phenylmethylsulfonyl fluoride. The cysteine proteinase hydrolyzes storage proteins of bombyx eggs but it was inactive with respect to N-benzoyl-D,L-arginine-p-nitroanilide (BAPNA). It is a cathepsin L-like enzyme.

摘要

通过色谱法和等电聚焦法纯化了在pH 3.0时具有最大活性的家蚕蛾卵半胱氨酸蛋白酶。在聚丙烯酰胺凝胶上进行SDS电泳时，纯化后的酶呈现出一条分子量为50 kD的单带。等电聚焦显示，家蚕蛾卵半胱氨酸蛋白酶以两种形式存在，其pI值分别为5.95和6.43。该酶的活性对碘乙酰胺和对氯汞苯甲酸的抑制敏感，但对EDTA、胃蛋白酶抑制剂和苯甲基磺酰氟具有抗性。这种半胱氨酸蛋白酶可水解家蚕蛾卵的储存蛋白，但对N-苯甲酰-D,L-精氨酸对硝基苯胺（BAPNA）无活性。它是一种组织蛋白酶L样酶。