The functional role of cytochrome b5 reincorporated into hepatic microsomal fractions.
Incorporation of detergent-solubilized cytochrome b5 into phenobarbital-induced rabbit liver microsomal fractions decelerates hexobarbital-dependent reduction of ferric cytochrome P-450; this is accompanied by retardation of NADPH utilization and H2O2 formation in the assay media. Integration of manganese-substituted cytochrome b5 into the microsomal preparations fails to affect these parameters. Analysis of the cytochrome P-450 reduction kinetics in the presence of increasing amounts of cytochrome b5 reveals a gradual augmentation of the amplitude of slow-phase electron transfer at the expense of the relative contribution of the fast phase; finally, a slow, apparently monophasic reaction persists. This defect in enzymatic reduction is not due to detergent effects and also does not seem to reflect cytochrome b5-induced perturbation of anchoring of NADPH-cytochrome c(P-450) reductase to cytochrome P-450. Experiments with the highly purified cytochrome P-450 isozyme LM2, in which amino acid residue(s) close to the heme edge had undergone suicidal inactivation through covalent attachment of chloramphenicol metabolite(s) do not exclude the possibility that cytochrome b5 and reductase might compete for a common electron transmission site on the terminal acceptor. Hence, the inhibitory action of cytochrome b5 on the reduction of ferric cytochrome P-450 is tentatively attributed to partial substitution of the former pigment for reductase in direct transport of the first electron to the monooxygenase.
D000818:Animals; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis / Q000494:pharmacology; D003573:Cytochrome b Group / Q000378:metabolism / Q000502:physiology*; D015786:Cytochromes b5; D004347:Drug Interactions; D004790:Enzyme Induction / Q000187:drug effects; D008862:Microsomes, Liver / Q000378:metabolism*; D009249:NADP / Q000502:physiology; D009251:NADPH-Ferrihemoprotein Reductase / Q000494:pharmacology; D011817:Rabbits
D003573:Cytochrome b Group; D009249:NADP; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase