Differences in the mechanism of functional interaction between NADPH-cytochrome P-450 reductase and its redox partners.
Selective methylamidation of NADPH-cytochrome P-450 reductase (EC 1.6.2.4) carboxyl groups was used to assess the relative importance of these groups in the enzyme-catalyzed reduction of cytochromes c, b5, and P-450. Methylamidation of as few as 7 mol of carboxyl groups per mol of reductase caused 80% inhibition of cytochrome c reduction, 50% inhibition of rat liver microsomal RLM3 reduction, and up to 90% inhibition in the capacity of the reductase to support reconstituted monooxygenase activities of RLM3, RLM5, and LM2. In marked contrast, cytochrome b5 reduction measured under comparable conditions was stimulated by 50%. The impaired interactions between the reductase and cytochromes P-450 LM2 and RLM5 were shown not to arise from an impaired capacity for the proteins to bind each other but more likely to be due to an inhibition of a step(s) subsequent to complex formation between the oxidized proteins. These results show that the reductase interacts functionally with cytochrome c and cytochromes P-450 on the one hand and cytochrome b5 on the other through different mechanisms.
D000818:Animals; D002021:Buffers; D055598:Chemical Phenomena; D002621:Chemistry; D003374:Coumarins / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003573:Cytochrome b Group / Q000378:metabolism; D003574:Cytochrome c Group / Q000378:metabolism; D015786:Cytochromes b5; D004579:Electron Transport; D007096:Imidoesters; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D008744:Methylamines; D008862:Microsomes, Liver / Q000201:enzymology; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism*; D010084:Oxidation-Reduction; D011817:Rabbits; D051381:Rats; D013329:Structure-Activity Relationship
D002021:Buffers; D003374:Coumarins; D003573:Cytochrome b Group; D003574:Cytochrome c Group; D007096:Imidoesters; D007527:Isoenzymes; D008744:Methylamines; C014539:ethyl acetimidate; C017299:7-ethoxycoumarin; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; C027451:methylamine; D009251:NADPH-Ferrihemoprotein Reductase