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

FEBS Letters

Gelhaye, Eric¹  Jacquot, Jean-Pierre¹  Rouhier, Nicolas¹ 

[1] Unité Mixte de Recherches INRA, Université Henri-Poincaré, UMR IaM 1136, Faculté des Sciences, P.O. Box 239, 54506 Vandoeuve Cedex, France

关键词: Glutaredoxin;    Thioredoxin;    Redox regulation;    Dithiol/disulfide exchange;    DHA;    dehydroascorbate;    DTT;    dithiothreitol;    Grx;    glutaredoxin;    GR;    glutathione reductase;    HED;    2-hydroxyethyldisulfide;    NADP-MDH;    NADP-malate dehydrogenase;    Prx;    peroxiredoxin;    Trx;    thioredoxin;   

DOI  :  10.1016/S0014-5793(01)03302-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 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.

【 授权许可】

Unknown