ACTIVE-SITE RESIDUES IN M-CALPAIN - IDENTIFICATION BY SITE-DIRECTED MUTAGENESIS

被引：52

作者：

ARTHUR, JSC ^{[1
]}

GAUTHIER, S ^{[1
]}

ELCE, JS ^{[1
]}

机构：

[1] QUEENS UNIV,DEPT BIOCHEM,KINGSTON,ON K7L 3N6,CANADA

来源：

FEBS LETTERS | 1995年 / 368卷 / 03期

基金：

英国医学研究理事会;

关键词：

CALPAIN; CA2+-BINDING; CATALYTIC TRIAD; CYSTEINE PROTEINASE; MUTAGENESIS;

D O I：

10.1016/0014-5793(95)00691-2

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Site-directed mutagenesis was used to alter putative active site residues in the large subunit of calpain, and the activity of the mutants was measured follo,wing coexpression in E. coli of both calpain subunits and purification of the resultant dimers, Mutants Cys105Ser, His262Ala and Asn286Ala had no activity. Together with sequence comparisons among cysteine proteinases, the results suggest that these residues constitute the catalytic triad in calpain. Mutants Asn286Asp and Trp288Tyr had low activity, consistent with interaction of these residues with His262.

引用

页码：397 / 400

页数：4

共 22 条

[1] ALIGNMENT PHYLOGENY OF THE PAPAIN SUPERFAMILY OF CYSTEINE PROTEASES [J].