title: Molecular docking and NMR binding studies to identify novel inhibitors of human phosphomevalonate kinase.
abstract: Phosphomevalonate kinase (PMK) phosphorylates mevalonate-5-phosphate (M5P) in the mevalonate pathway, which is the sole source of isoprenoids and steroids in humans. We have identified new PMK inhibitors with virtual screening, using autodock. Promising hits were verified and their affinity measured using NMR-based (1)H-(15)N heteronuclear single quantum coherence (HSQC) chemical shift perturbation and fluorescence titrations. Chemical shift changes were monitored, plotted, and fitted to obtain dissociation constants (K(d)). Tight binding compounds with K(d)'s ranging from 6-60 μM were identified. These compounds tended to have significant polarity and negative charge, similar to the natural substrates (M5P and ATP). HSQC cross peak changes suggest that binding induces a global conformational change, such as domain closure. Compounds identified in this study serve as chemical genetic probes of human PMK, to explore pharmacology of the mevalonate pathway, as well as starting points for further drug development.
authors.affiliation: Chemical Proteomics Facility at Marquette, Department of Chemistry, Marquette University, Milwaukee, WI 53201, United States.
authors.forename: Pornthip
authors.initials: P
authors.lastname: Boonsri
authors.forename: Terrence S
authors.initials: TS
authors.lastname: Neumann
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Olson
authors.forename: Sheng
authors.initials: S
authors.lastname: Cai
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Herdendorf
authors.forename: Henry M
authors.initials: HM
authors.lastname: Miziorko
authors.forename: Supa
authors.initials: S
authors.lastname: Hannongbua
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Sem
mesh_terms: D020134:Catalytic Domain / Q000187:drug effects; D018360:Crystallography, X-Ray; D055808:Drug Discovery / Q000379:methods*; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000037:antagonists & inhibitors* / Q000737:chemistry; D017433:Protein Structure, Secondary
chemical_list: D004791:Enzyme Inhibitors; D017856:Phosphotransferases (Phosphate Group Acceptor); C022371:phosphomevalonate kinase