Surface-modified mutants of cytochrome P450cam: enzymatic properties and electrochemistry.
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.
Lo
K K
KK
Department of Chemistry, Inorganic Chemistry Laboratory, Oxford, UK.
Wong
L L
LL
Hill
H A
HA
D000818:Animals; D019475:Camphor 5-Monooxygenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004789:Enzyme Activation / Q000235:genetics; D004926:Escherichia coli; D009154:Mutation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D055672:Static Electricity
D011994:Recombinant Proteins; D019475:Camphor 5-Monooxygenase