Exploring the active site of plant glutaredoxin by site-directed mutagenesis

被引：60
|
作者：
Rouhier, N [1 ]
Gelhaye, E [1 ]
Jacquot, JP [1 ]
机构：
[1] Univ Henri Poincare, UMR INRA, UMR IaM 1136, Fac Sci, F-54506 Vandoeuvre Les Nancy, France
来源：
FEBS LETTERS | 2002年 / 511卷 / 1-3期
关键词：
glutaredoxin; thioredoxin; redox regulation; dithiol/disulfide exchange;
D O I：
10.1016/S0014-5793(01)03302-6
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：
Six mutants (Y26A, C27S, Y29F, Y29P, C30S and Y26W/Y29P) have been engineered in order to explore the active site of poplar glutaredoxin (Grx) (Y26CPYC30). The cysteinic mutants indicate that Cys 27 is the primary nucleophile. Phe is a good substitute for Tyr 29, but the Y29P mutant was inactive. The Y26A mutation caused a moderate loss of activity. The YCPPC and WCPPC mutations did not improve the reactivity of Grx with the chloroplastic NADP-malate dehydrogenase, a well known target of thioredoxins (Trxs). The results are discussed in relation with the known biochemical properties of Grx and Trx. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码：145 / 149
页数：5
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