title: A structure-based approach to ligand discovery for 2C-methyl-D-erythritol-2,4-cyclodiphosphate synthase: a target for antimicrobial therapy.
abstract: The nonmevalonate route to isoprenoid biosynthesis is essential in Gram-negative bacteria and apicomplexan parasites. The enzymes of this pathway are absent from mammals, contributing to their appeal as chemotherapeutic targets. One enzyme, 2C-methyl-d-erythritol-2,4-cyclodiphosphate synthase (IspF), has been validated as a target by genetic approaches in bacteria. Virtual screening against Escherichia coli IspF (EcIspF) was performed by combining a hierarchical filtering methodology with molecular docking. Docked compounds were inspected and 10 selected for experimental validation. A surface plasmon resonance assay was developed and two weak ligands identified. Crystal structures of EcIspF complexes were determined to support rational ligand development. Cytosine analogues and Zn(2+)-binding moieties were characterized. One of the putative Zn(2+)-binding compounds gave the lowest measured K(D) to date (1.92 +/- 0.18 muM). These data provide a framework for the development of IspF inhibitors to generate lead compounds of therapeutic potential against microbial pathogens.
authors.affiliation: Division of Biological Chemistry and Drug Discovery, College of Life Sciences, University of Dundee, Dow Street, Dundee, DD1 5EH, Scotland, United Kingdom.
authors.forename: Nicola L
authors.initials: NL
authors.lastname: Ramsden
authors.forename: Lori
authors.initials: L
authors.lastname: Buetow
authors.forename: Alice
authors.initials: A
authors.lastname: Dawson
authors.forename: Lauris A
authors.initials: LA
authors.lastname: Kemp
authors.forename: Venkatsubramanian
authors.initials: V
authors.lastname: Ulaganathan
authors.forename: Ruth
authors.initials: R
authors.lastname: Brenk
authors.forename: Gerhard
authors.initials: G
authors.lastname: Klebe
authors.forename: William N
authors.initials: WN
authors.lastname: Hunter