Surface-modified mutants of cytochrome P450cam: enzymatic properties and electrochemistry.

Journal article

We report the electrochemistry of genetic variants of the haem monooxygenase cytochrome P450cam. 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.

Authors: Lo, K; Wong, L; Hill, H

• Publication status: Published
• Journal: FEBS letters
• Volume: 451
• Issue: 3
• Pages: 342-346
• Publication date: 1999-05-01
• DOI: 10.1016/s0014-5793(99)00611-0
• EISSN: 1873-3468
• ISSN: 0014-5793
• Language: English
• Keywords: Enzyme Activation; Mutation; Static Electricity; Animals; Camphor 5-Monooxygenase; Escherichia coli; Recombinant Proteins