Stoichiometry of electrocatalytic cycle of cytochrome P450 2B4.
Stoichiometry of the electrocatalytical cycle of cytochrome P450 2B4 was studied in kinetic mode according to bielectrode scheme. Graphite screen-printed electrodes with immobilized cytochrome P450 2B4 were used as the operating electrode (at the potential E(0')=-450 mV) and electrodes, modified with cytochrome c (E(0')=-50 mV) or Prussian Blue (E(0')=0), as measuring electrodes (for H(2)O(2)) and Clark-type electrode (for O(2)). Benzphetamine N-demethylation rate was 17+/-3 nmol/nmol of enzyme/min, peroxide production was 4.8+/-0.7 nmol/nmol of enzyme/min (substrate-free system), 3.3+/-0.6 nmol/nmol of enzyme/min (0.5 mM benzphetamine), the oxygen consumption rate by capital P450 2B4 was 19.4+/-0.6 nmol/nmol of enzyme/min (in the presence of benzphetamine), 4.8+/-0.4 nmol/nmol of enzyme/min (without substrate). Based on stoichiometry of P450 electrocatalysis adequacy of electrochemical reduction and P450-monooxygenase system was revealed.
Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Pogodinskaya Street, 10, Moscow 119121, Russia.
Yuri O
YO
Rudakov
Victoria V
VV
Shumyantseva
Tatiana V
TV
Bulko
Elena V
EV
Suprun
Galina P
GP
Kuznetsova
Natalia F
NF
Samenkova
Alexander I
AI
Archakov
D001189:Aryl Hydrocarbon Hydroxylases / Q000737:chemistry*; D002384:Catalysis; D000072467:Cytochrome P450 Family 2; D004563:Electrochemistry; D006801:Humans; D007700:Kinetics; D053758:Nanoparticles; D010084:Oxidation-Reduction
D001189:Aryl Hydrocarbon Hydroxylases; D000072467:Cytochrome P450 Family 2; C465241:cytochrome P-450 CYP2B4 (rabbit)