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Surface‐modified mutants of cytochrome P450 cam : enzymatic properties and electrochemistry
Author(s) -
Lo Kenneth Kam-Wing,
Wong Luet-Lok,
Hill H.Allen O
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)00611-0
Subject(s) - cysteine , chemistry , mutant , electrochemistry , hydroxylation , monooxygenase , cytochrome , stereochemistry , cytochrome p450 , biochemistry , enzyme , electrode , gene
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.