Proton nuclear magnetic resonance study of the binary complex of cytochrome P450cam and putidaredoxin: interaction and electron transfer rate analysis.
A 1H nuclear magnetic resonance study of the complex of cytochrome P450cam-putidaredoxin has been performed. Isocyanide is bound to cytochrome P450cam in order to increase the stability of the protein both in the reduced and the oxidized state. Diprotein complex formation was detected through variation of the heme methyl proton resonances which have been assigned in the two redox states. The electron transfer rate at equilibrium was determinated by magnetization transfer experiments. The observed rate of oxidation of reduced cytochrome P450 by the oxidized putidaredoxin is 27 (+/- 7) per s.
Mouro
C
C
Laboratoire de Chimie Organométallique et Biologique, UMR CNRS 6509, Université de Rennes 1, France.
Bondon
A
A
Jung
C
C
Hui Bon Hoa
G
G
De Certaines
J D
JD
Spencer
R G
RG
Simonneaux
G
G
D019475:Camphor 5-Monooxygenase / Q000737:chemistry*; D004579:Electron Transport; D004795:Enzyme Stability; D005288:Ferredoxins / Q000737:chemistry*; D006418:Heme / Q000737:chemistry; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy; D009570:Nitriles / Q000737:chemistry; D011522:Protons
D005288:Ferredoxins; D046911:Macromolecular Substances; D009570:Nitriles; D011522:Protons; C002990:butyl isocyanide; D006418:Heme; C100137:putidaredoxin; D019475:Camphor 5-Monooxygenase