Surface‐modified mutants of cytochrome P450cam: enzymatic properties and electrochemistry
- 28 May 1999
- journal article
- Published by Wiley in FEBS Letters
- Vol. 451 (3) , 342-346
- https://doi.org/10.1016/s0014-5793(99)00611-0
Abstract
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.Keywords
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