title: Structural basis for cooperative binding of azoles to CYP2E1 as interpreted through guided molecular dynamics simulations.
abstract: CYP2E1 metabolizes a wide array of small, hydrophobic molecules, resulting in their detoxification or activation into carcinogens through Michaelis-Menten as well as cooperative mechanisms. Nevertheless, the molecular determinants for CYP2E1 specificity and metabolic efficiency toward these compounds are still unknown. Herein, we employed computational docking studies coupled to molecular dynamics simulations to provide a critical perspective for understanding a structural basis for cooperativity observed for an array of azoles from our previous binding and catalytic studies (Hartman et al., 2014). The resulting 28 CYP2E1 complexes in this study revealed a common passageway for azoles that included a hydrophobic steric barrier causing a pause in movement toward the active site. The entrance to the active site acted like a second sieve to restrict access to the inner chamber. Collectively, these interactions impacted the final orientation of azoles reaching the active site and hence could explain differences in their biochemical properties observed in our previous studies, such as the consequences of methylation at position 5 of the azole ring. The association of a second azole demonstrated significant differences in interactions stabilizing the bound complex than observed for the first binding event. Intermolecular interactions occurred between the two azoles as well as CYP2E1 residue side chains and backbone and involved both hydrophobic contacts and hydrogen bonds. The relative importance of these interactions depended on the structure of the respective azoles indicating the absence of specific defining criteria for binding unlike the well-characterized dominant role of hydrophobicity in active site binding. Consequently, the structure activity relationships described here and elsewhere are necessary to more accurately identify factors impacting the observation and significance of cooperativity in CYP2E1 binding and catalysis toward drugs, dietary compounds, and pollutants.
authors.affiliation: Department of Chemistry and Physics, Ouachita Baptist University, Arkadelphia, AR, USA.
authors.forename: Joseph W
authors.initials: JW
authors.lastname: Levy
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA.
authors.forename: Jessica H
authors.initials: JH
authors.lastname: Hartman
authors.affiliation: Department of Chemistry and Physics, Ouachita Baptist University, Arkadelphia, AR, USA.
authors.forename: Martin D
authors.initials: MD
authors.lastname: Perry
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, AR, USA. Electronic address: millergroverp@uams.edu.
authors.forename: Grover P
authors.initials: GP
authors.lastname: Miller
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001393:Azoles / Q000737:chemistry*; D020134:Catalytic Domain; D019392:Cytochrome P-450 CYP2E1 / Q000737:chemistry*; D065607:Cytochrome P-450 Enzyme Inhibitors / Q000737:chemistry*; D015195:Drug Design; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008024:Ligands; D062105:Molecular Docking Simulation; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D011485:Protein Binding; D011817:Rabbits; D055672:Static Electricity; D013329:Structure-Activity Relationship; D013816:Thermodynamics
chemical_list: D001393:Azoles; D065607:Cytochrome P-450 Enzyme Inhibitors; D008024:Ligands; D019392:Cytochrome P-450 CYP2E1
keywords: Allostery; Azole; Cytochrome P450; Docking; Molecular dynamics; Structure activity relationships