NADPH-P-450 reductase: Structural and functional comparisons of the eukaryotic and prokaryotic isoforms

Abstract

The comparison of the properties of microsomal NADPH-P-450 reductase and the flavoprotein domain of P-450 BM-3 (BMR) has revealed a significant difference in the mechanism of reduction of the hemoprotein P-450 by these flavoproteins. Microsomal NADPH-P-450 reductase transfers electrons to the hemoprotein by shuttling between hydroquinone and semiquinone forms of the FMN delivering one electron per cycle. Since the microsomal NADPH-P450 reductase has evolved as a component of multienzyme system, this type of mechanism may permit regulation of the steps of the P-450 reaction via variation in the affinity of the reductase for different P-450s, interaction with cytochrome b 5, etc. In contrast, in the soluble, bacterial flavocytochrome P-450 BM-3, the reductase domain has evolved together with a single unique heme domain. This enzyme was found to utilize the fastest and simplest way to reduce the heme iron, with the FMN moiety of BMR shuttling between the semiquinone and oxidized states. This mechanism of reduction provides the highest turnover number of any P-450 and tight coupling of the monooxygenation reaction. While there are clear differences in the intermediates involved in the reduction of P-450s by these two enzymes, the domain structure and presumably the mode of interaction between the reductase and P-450s has been maintained over evolutionary time.