title: Construction of 3D models of the CYP11B family as a tool to predict ligand binding characteristics.
abstract: Aldosterone is synthesised by aldosterone synthase (CYP11B2). CYP11B2 has a highly homologous isoform, steroid 11beta-hydroxylase (CYP11B1), which is responsible for the biosynthesis of aldosterone precursors and glucocorticoids. To investigate aldosterone biosynthesis and facilitate the search for selective CYP11B2 inhibitors, we constructed three-dimensional models for CYP11B1 and CYP11B2 for both human and rat. The models were constructed based on the crystal structure of Pseudomonas Putida CYP101 and Oryctolagus Cuniculus CYP2C5. Small steric active site differences between the isoforms were found to be the most important determinants for the regioselective steroid synthesis. A possible explanation for these steric differences for the selective synthesis of aldosterone by CYP11B2 is presented. The activities of the known CYP11B inhibitors metyrapone, R-etomidate, R-fadrazole and S-fadrazole were determined using assays of V79MZ cells that express human CYP11B1 and CYP11B2, respectively. By investigating the inhibitors in the human CYP11B models using molecular docking and molecular dynamics simulations we were able to predict a similar trend in potency for the inhibitors as found in the in vitro assays. Importantly, based on the docking and dynamics simulations it is possible to understand the enantioselectivity of the human enzymes for the inhibitor fadrazole, the R-enantiomer being selective for CYP11B2 and the S-enantiomer being selective for CYP11B1.
authors.affiliation: BioModeling and BioInformatics, Eindhoven University of Technology, Eindhoven, The Netherlands. L.Roumen@tue.nl
authors.forename: Luc
authors.initials: L
authors.lastname: Roumen
authors.forename: Marijn P A
authors.initials: MP
authors.lastname: Sanders
authors.forename: Koen
authors.initials: K
authors.lastname: Pieterse
authors.forename: Peter A J
authors.initials: PA
authors.lastname: Hilbers
authors.forename: Ralf
authors.initials: R
authors.lastname: Plate
authors.forename: Erica
authors.initials: E
authors.lastname: Custers
authors.forename: Marcel
authors.initials: M
authors.lastname: de Gooyer
authors.forename: Jos F M
authors.initials: JF
authors.lastname: Smits
authors.forename: Ilona
authors.initials: I
authors.lastname: Beugels
authors.forename: Judith
authors.initials: J
authors.lastname: Emmen
authors.forename: Harry C J
authors.initials: HC
authors.lastname: Ottenheijm
authors.forename: Dirk
authors.initials: D
authors.lastname: Leysen
authors.forename: J J R
authors.initials: JJ
authors.lastname: Hermans
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D020134:Catalytic Domain / Q000235:genetics; D003198:Computer Simulation; D019405:Cytochrome P-450 CYP11B2 / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015195:Drug Design; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D051381:Rats; D017386:Sequence Homology, Amino Acid; D013252:Steroid 11-beta-Hydroxylase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013816:Thermodynamics
chemical_list: D008024:Ligands; D019405:Cytochrome P-450 CYP11B2; D013252:Steroid 11-beta-Hydroxylase