Structural and functional effects of cytochrome b₅ interactions with human cytochrome P450 enzymes

The small heme-containing protein cytochrome b₅ can facilitate, inhibit, or have no effect on cytochrome P450 catalysis, often in a P450-dependent and substrate-dependent manner that is not well understood. Herein, solution NMR was used to identify b₅ residues interacting with different human drug-metabolizing P450 enzymes. NMR results revealed that P450 enzymes bound to either b₅ α4-5 (CYP2A6 and CYP2E1) or this region and α2-3 (CYP2D6 and CYP3A4) and suggested variation in the affinity for b₅ Mutations of key b₅ residues suggest not only that different b₅ surfaces are responsible for binding different P450 enzymes, but that these different complexes are relevant to the observed effects on P450 catalysis.