Surface‐modified mutants of cytochrome P450cam: enzymatic properties and electrochemistry

FEBS Letters

Wong, Luet-Lok¹  Hill, H.Allen O¹  Lo, Kenneth Kam-Wing¹ 

[1] Department of Chemistry, Inorganic Chemistry Laboratory, South Parks Road, Oxford OX1 3QR, UK

关键词: Electrochemistry;    Mutagenesis;    P450cam;    Redox-active label;    Surface cysteine;   

DOI  :  10.1016/S0014-5793(99)00611-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We report the electrochemistry of genetic variants of the haem monooxygenase cytochrome P450_cam. A surface cysteine-free mutant (abbreviated as SCF) was prepared in which the five surface cysteine residues Cys-58, Cys-85, Cys-136, Cys-148 and Cys-334 were changed to alanines. Four single surface cysteine mutants with an additional mutation, R72C, R112C, K344C or R364C, were also prepared. The haem spin-state equilibria, NADH turnover rates and camphor-hydroxylation properties, as well as the electrochemistry of these mutants are reported. The coupling of a redox-active label, N-ferrocenylmaleimide, to the single surface cysteine mutant SCF-K344C, and the electrochemistry of this modified mutant are also described.

【 授权许可】

Unknown