title: A novel allosteric inhibitor of the uridine diphosphate N-acetylglucosamine pyrophosphorylase from Trypanosoma brucei.
abstract: Uridine diphosphate N-acetylglucosamine pyrophosphorylase (UAP) catalyzes the final reaction in the biosynthesis of UDP-GlcNAc, an essential metabolite in many organisms including Trypanosoma brucei, the etiological agent of Human African Trypanosomiasis. High-throughput screening of recombinant T. brucei UAP identified a UTP-competitive inhibitor with selectivity over the human counterpart despite the high level of conservation of active site residues. Biophysical characterization of the UAP enzyme kinetics revealed that the human and trypanosome enzymes both display a strictly ordered bi-bi mechanism, but with the order of substrate binding reversed. Structural characterization of the T. brucei UAP-inhibitor complex revealed that the inhibitor binds at an allosteric site absent in the human homologue that prevents the conformational rearrangement required to bind UTP. The identification of a selective inhibitory allosteric binding site in the parasite enzyme has therapeutic potential.
authors.affiliation: Division of Biological Chemistry and Drug Discovery, ‡Division of Molecular Microbiology, and §MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee , Dundee DD1 5EH, U.K.
authors.forename: Michael D
authors.initials: MD
authors.lastname: Urbaniak
authors.forename: Iain T
authors.initials: IT
authors.lastname: Collie
authors.forename: Wenxia
authors.initials: W
authors.lastname: Fang
authors.forename: Tonia
authors.initials: T
authors.lastname: Aristotelous
authors.forename: Susanne
authors.initials: S
authors.lastname: Eskilsson
authors.forename: Olawale G
authors.initials: OG
authors.lastname: Raimi
authors.forename: Justin
authors.initials: J
authors.lastname: Harrison
authors.forename: Iva Hopkins
authors.initials: IH
authors.lastname: Navratilova
authors.forename: Julie A
authors.initials: JA
authors.lastname: Frearson
authors.forename: Daan M F
authors.initials: DM
authors.lastname: van Aalten
authors.forename: Michael A J
authors.initials: MA
authors.lastname: Ferguson
mesh_terms: D000494:Allosteric Regulation / Q000187:drug effects; D020134:Catalytic Domain; D006801:Humans; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D014344:Trypanocidal Agents / Q000737:chemistry* / Q000494:pharmacology*; D014346:Trypanosoma brucei brucei / Q000187:drug effects* / Q000201:enzymology* / Q000254:growth & development; D014353:Trypanosomiasis, African / Q000188:drug therapy / Q000469:parasitology; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000378:metabolism
chemical_list: D014344:Trypanocidal Agents; D014537:Uridine Diphosphate N-Acetylglucosamine; D009713:Nucleotidyltransferases; C023226:UDPacetylglucosamine pyrophosphorylase