The role of CYP3A-CYP2E1 interactions in activation of CYP3A enzymes by chronic alcohol exposure

Abstract Aiming to examine the effect of chronic alcohol exposure on the activity of CYP3A enzymes in human liver, we studied the metabolism of two CYP3A-specific substrates, 7-benzyloxyquinoline (7-BQ) and ivermectin, in 23 preparations of human liver microsomes (HLM) obtained from donors with documented alcohol exposure, graded from non-drinkers to heavy alcoholics. All HLM samples were characterized for the composition of the cytochrome P450 pool and the abundances of other drug-metabolizing and endoplasmic reticulum-stress-related enzymes by global proteomics. Our studies revealed a striking increase in the activities of CYP3A enzymes caused by chronic alcohol exposure. This effect is not associated with CYP3A enzyme levels, which do not correlate with alcohol exposure. Instead, the rates of 7-BQ and ivermectin metabolism correlate with the content of alcohol-inducible CYP2E1. However, this enzyme does not metabolize ivermectin, and its activity with 7-BQ is negligible. These results suggest that the observed acceleration of the elimination of drugs metabolized by CYP3A enzymes by alcohol exposure is due to functional effects of the interaction between CYP3A and CYP2E1. To elucidate the potential mechanism of this effect, we studied the formation of CYP2E1-CYP3A4 complexes in CYP3A4-containing Supersomes with co-incorporated CYP2E1 using tag-transfer chemical crosslinking mass spectrometry (CX-MS). These experiments confirmed physical interactions between the proteins and allowed the identification of CYP3A4 residues at the sites of contact. This information was used to build structural models of the CYP2E1-CYP3A4 complex and to propose possible mechanisms for the observed effects. Competing Interest Statement All authors except BP declare no competing interest for this work. BP is a cofounder of Precision Quantomics Inc. and recipient of research funding from Bristol Myers Squibb, Genentech, Gilead, Merck, Novartis, Takeda, and Generation Bio. Footnotes We have corrected quite a few typos and edited several sentences to bring this version into alignment with the one we plan to submit to the journal.