Characteristics of microsomal reduction of benzo[a]pyrene 4,5-oxide.
NADPH-reduction of benzo[a]pyrene 4,5-oxide (BP-4,5-oxide) to BP required four components from rat liver: cytochrome P-450, NADPH cytochrome P-450 reductase, phosphatidylcholine and a soluble, heat-sensitive factor which was present in 105 000 X g supernatant and was also released from microsomes by sonication. The requirement for this factor contrasts with recently reported results from Sugiura et al. (Cancer Res., 40 (1980) 2910). Oxide-reduction was 40 times faster under anaerobic conditions, but oxygen did not affect the stimulation factor. This stimulation was highest (X15) at low concentrations of microsomal protein (less than 0.1 mg/ml) and was almost absent at high concentrations of microsomal protein (greater than 1 mg/ml). Oxide-reduction activity was proportional to microsomal protein concentration in the presence of added 105 000 X g supernatant, but for microsomes alone (greater than 0.1 mg/ml) exhibited a parallel plot with an intercept at 0.08 mg/ml microsomal protein. Stimulation was highest at high concentrations of BP-4,5-oxide and a linear plot of V-1 vs. [BP-4,5-oxide]-1 was only obtained in the presence of 105 000 X g supernatant (Km = 3 microM, Vmax = 3.3 nmol/mg/min). Microsomal hydration of BP-4,5-oxide (inhibited in reductase assays) was unaffected by 105 000 X g supernatant, suggesting that stimulation of oxide-reduction did not derive from solubilization of BP-4,5-oxide. Stimulation was observed in the initial rate of reaction and was independent of incubation time. Inhibition of lipid peroxidation, removal of peroxides and deoxygenation were all excluded as explanations of the stimulatory effect.
D000693:Anaerobiosis; D000818:Animals; D001564:Benzo(a)pyrene; D001580:Benzopyrenes / Q000378:metabolism*; D002458:Cell Fractionation; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D008748:Methylcholanthrene / Q000494:pharmacology; D008862:Microsomes, Liver / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D010084:Oxidation-Reduction; D010634:Phenobarbital / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000378:metabolism; D051381:Rats
D001580:Benzopyrenes; D010713:Phosphatidylcholines; D001564:Benzo(a)pyrene; C006707:benzo(a)pyrene 4,5-epoxide; D008748:Methylcholanthrene; D003577:Cytochrome P-450 Enzyme System; D009251:NADPH-Ferrihemoprotein Reductase; D010634:Phenobarbital