Engineering of an intersubunit disulphide bridge in glutathione reductase from Escherichia coli

FEBS Letters

Scrutton, Nigel S.¹  Perham, Richard N.¹  Berry, Alan¹ 

[1] Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge CB2 1QW, England

关键词: Glutathione reductase;    Protein engineering;    Site-directed mutagenesis;    Disulfide bridge;    Thermal stability;    NADPH;   

DOI  :  10.1016/0014-5793(88)81028-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

By site-directed mutagenesis, Thr-75 was converted to Cys-75 in the glutathione reductase (EC 1.6.4.2 of Escherichia coli. This led to the spontaneous formation of an intersubunit disulphide bridge across the 2-fold axis of the dimeric enzyme. The disulphide bridge had no deleterious effect on the catalytic activity, but nor did it increase the thermal stability of the enzyme, possibly because of local conformational flexibility on the dimer interface. The T75C mutant, like the wild-type enzyme, was inactivated by NADPH, proving that this inactivation cannot be due to simple dissociation of the dimer.

【 授权许可】

Unknown