title: QM/MM modeling of benzene hydroxylation in human cytochrome P450 2C9.
abstract: The mechanism of benzene hydroxylation was investigated in the realistic enzyme environment of the human CYP 2C9 by using quantum mechanical/molecular mechanical (QM/MM) calculations of the whole reaction profile using the B3LYP method to describe the QM region. The calculated QM/MM barriers for addition of the active species Compound I to benzene are consistent with experimental rate constants for benzene metabolism in CYP 2E1. In contrast to gas-phase model calculations, our results suggest that competing side-on and face-on geometries of arene addition may both occur in the case of aromatic ring oxidation in cytochrome P450s. QM/MM profiles for three different rearrangement pathways of the initially formed sigma-adduct, leading to formation of epoxide, ketone, and an N-protonated porphyrin species, were calculated. Our results suggest that epoxide and ketone products form with comparable ease in the face-on pathway, whereas epoxide formation is preferred in the side-on pathway. Additionally, rearrangement to the N-protonated porphyrin species was found to be competitive with side-on epoxide formation. This suggests that overall, the competition between formation of epoxide and phenol final products in P450 oxidation of aromatic substrates is quite finely balanced.
authors.affiliation: Centre for Computational Chemistry, School of Chemistry, University of Bristol, Cantocks' Close, Bristol BS8 1TS, UK.
authors.forename: Christine M
authors.initials: CM
authors.lastname: Bathelt
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Mulholland
authors.forename: Jeremy N
authors.initials: JN
authors.lastname: Harvey