The role of protein-protein and protein-membrane interactions on P450 function

This symposium summary, sponsored by the ASPET, was held at Experimental Biology 2015 on March 29, 2015, in Boston, Massachusetts. The symposium focused on: 1) the interactions of cytochrome P450s (P450s) with their redox partners; and 2) the role of the lipid membrane in their orientation and stabilization. Two presentations discussed the interactions of P450swithNADPH-P450 reductase (CPR) and cytochrome b₅. First, solution nuclear magnetic resonance was used to compare the protein interactions that facilitated either the hydroxylase or lyase activities of CYP17A1. The lyase interaction was stimulated by the presence of b5 and 17a-hydroxypregnenolone, whereas the hydroxylase reaction was predominant in the absence of b5. Theroleof b5 wasalso shownin vivo by selective hepatic knockout of b5 from mice expressing CYP3A4 and CYP2D6; the lack of b₅ caused a decrease in the clearance of several substrates. The role of the membrane on P450 orientation was examined using computational methods, showing that the proximal region of the P450 molecule faced the aqueous phase. The distal region, containing the substrate-Access channel, was associated with the membrane. The interaction of NADPH-P450 reductase (CPR) with themembrane was also described, showing the ability of CPR to "helicopter" above the membrane. Finally, the endoplasmic reticulum (ER) was shown to be heterogeneous, having ordered membrane regions containing cholesterol and more disordered regions. Interestingly, two closely related P450s, CYP1A1 and CYP1A2, resided in different regions of the ER. The structural characteristics of their localization were examined. These studies emphasize the importance ofP450 protein organization to their function.