Oxycytochrome P-450: its breakdown to superoxide for the formation of hydrogen peroxide.
Four different experimental studies are described which were designed to evaluate the role of oxycytochrome P-450 in the formation of superoxide anions and hydrogen peroxide. The use of lipophilic copper chelates with superoxide dismutase like activity revealed that the primary site of interaction of these agents is related to the inhibition of the flavoprotein. NADPH-cytochrome P-450 reductase. Measurements of the proton assisted nucleophilic displacement of superoxide from oxycytochrome P-450 by high concentrations of sodium azide indicated an increase in the rate of hydrogen peroxide formation concomitant with the inhibition of the N-demethylation of ethylmorphine. Studies on the effect of NADH on the rate of hydrogen peroxide formation during NADPH oxidation by liver microsomes failed to reveal a stimulatory or synergistic effect in a manner analogous to results obtained during the cytochrome P-450 dependent oxidation of substrates such as ethylmorphine. These results suggest that hydrogen peroxide formation may not require the reduction of oxycytochrome P-450 to peroxycytochrome P-450. Measurements of the reduction of succinylated cytochrome c using purified cytochrome P-450 and the flavoprotein, NADPH-cytochrome P-450 reductase, directly demonstrate the formation of superoxide anions. It is concluded that oxycytochrome P-450 may decompose to generate hydrogen peroxide.
D000818:Animals; D001386:Azides / Q000494:pharmacology; D003300:Copper / Q000494:pharmacology; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D003574:Cytochrome c Group / Q000378:metabolism; D006861:Hydrogen Peroxide; D007521:Isocitrate Dehydrogenase / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism*; D009243:NAD; D010084:Oxidation-Reduction; D010100:Oxygen; D010634:Phenobarbital / Q000494:pharmacology; D051381:Rats; D013481:Superoxides; D014443:Tyrosine / Q000494:pharmacology
D001386:Azides; D003574:Cytochrome c Group; D009243:NAD; D013481:Superoxides; D014443:Tyrosine; D003300:Copper; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; D007521:Isocitrate Dehydrogenase; D010100:Oxygen; D010634:Phenobarbital