Evidence for two ethanol oxidizing pathways in reconstituted mixed-function oxidase systems.
The oxidation of ethanol and typical hydroxyl radical scavengers by NADPH-cytochrome P-450 reductase and cytochrome P-450 purified from phenobarbital-treated rats were studied. Ethanol and the scavengers could be oxidized by the reductase system itself. This system was inhibited by superoxide dismutase, competing hydroxyl radical scavengers and desferrioxamine, but stimulated by either EDTA or iron. These results suggest that an iron-catalyzed Haber-Weiss reaction might be involved in the mechanism by which the reductase mediates the oxidation of typical hydroxyl radical scavengers and ethanol. The addition of cytochrome P-450 had no effect on the oxidation of the scavengers, whereas the oxidation of ethanol was enhanced two-to three-fold over the reductase-dependent rate. The oxidation of ethanol was dependent on both the amount of reductase and P-450. There was no effect of competing scavengers, superoxide dismutase or desferrioxamine on the increased rate of ethanol oxidation produced upon addition of cytochrome P-450. Organic hydroperoxides supported the oxidation of ethanol, but not that of the scavengers when added directly to cytochrome P-450. These results suggest that two independent pathways are operative in supporting NADPH-dependent microsomal oxidation of ethanol. One pathway involves hydroxyl radicals which can be generated by the reductase, whereas the other pathway requires the combined presence of both the reductase and cytochrome P-450, and appears to be independent of oxygen radicals.
D000818:Animals; D002374:Catalase / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D000431:Ethanol / Q000378:metabolism*; D006878:Hydroxides / Q000378:metabolism; D017665:Hydroxyl Radical; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology*; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D010084:Oxidation-Reduction; D051381:Rats; D013482:Superoxide Dismutase / Q000378:metabolism
D006878:Hydroxides; D017665:Hydroxyl Radical; D000431:Ethanol; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D002374:Catalase; D013482:Superoxide Dismutase; D009251:NADPH-Ferrihemoprotein Reductase