Allosteric Interactions in Human Cytochrome P450 CYP3A4: The Role of Phenylalanine 213

Abstract

The role of Phe213 in the allosteric mechanism of human cytochrome P450 CYP3A4 was studied using a combination of progesterone (PGS) and carbamazepine (CBZ) as probe substrates. We expressed, purified and incorporated in POPC Nanodiscs three mutants, F213A, F213S, and F213Y, and compared them with the wild-type CYP3A4 monitoring spectral titration, the rate of NADPH oxidation and steady-state product turnover rates with pure substrates and substrate mixtures. All mutants demonstrated higher activity with CBZ, lower activity with PGS, and reduced activation of CBZ epoxidation by PGS, most pronounced in F213A mutant. Using allatom molecular dynamics simulations we compared dynamics of WT CYP3A4 and the F213A mutant incorporated in the lipid bilayer and the effect of the presence of PGS molecule at the allosteric peripheral site and evaluated the critical role of Phe213 in mediating the heterotropic allosteric interactions in CYP3A4.

Mechanisms of allosteric and mixed mode aromatase inhibitors

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In vitro inhibitory effect of lysionotin on the activity of cytochrome P450 enzymes

“…CYP3A4 is a major member of the CYP3A family, which is involved in the liver and intestine metabolism of more than 40% of marketed drugs and pharmaceuticals and catalyses the transformation of various substrates (Denisov et al 2019). The inhibition of CYP3A4 could directly inhibit the metabolism of its substrates, such as warfarin, puerarin, and oridonin, which is the main factor that induce drug-drug interaction (Liu et al 2019a(Liu et al , 2019bSong et al 2020).…”

Alcohol-Induced Increase in the Content of CYP2E1 in Human Liver Microsomes Causes a Multifold Activation of CYP3A4 and Attenuates its Cooperativity

“…More recent studies compellingly demonstrated the presence of a separate allosteric effector binding site and revealed its role in the instances of heterotropic activation of CYP3A4 by various effectors, including ANF (Davydov et al, 2012;Davydov et al, 2013;Polic and Auclair, 2017;Denisov et al, 2018;Marsch et al, 2018;Denisov et al, 2019;Ducharme et al, 2019). This allosteric site is formed at the vicinity of the F' and G' helices and located at the interface between the two CYP3A4 molecules in the crystallographic dimer of CYP3A4 complex with peripherally-bound progesterone (PDB 1W0F (Williams et al, 2004)).…”