Redox control of the P450cam catalytic cycle: effects of Y96F active site mutation and binding of a non-natural substrate.
Spectroelectrochemistry measurements are used to demonstrate that active site mutation and binding of an non-natural substrate to P450cam (CYP101) reduces the shift in the redox potential caused by substrate-binding, and thereby results in slower catalytic turnover rate relative to wild-type enzyme with the natural camphor substrate.
Reipa
Vytas
V
Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA. vytas@nist.gov
Mayhew
Martin P
MP
Holden
Marcia J
MJ
Vilker
Vincent L
VL
D001419:Bacteria / Q000201:enzymology / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D019475:Camphor 5-Monooxygenase / Q000378:metabolism*; D002384:Catalysis; D004563:Electrochemistry; D004579:Electron Transport; D009154:Mutation / Q000235:genetics; D009243:NAD / Q000737:chemistry; D010084:Oxidation-Reduction; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics
D009243:NAD; D019475:Camphor 5-Monooxygenase