TY - DATA T1 - A Novel Allosteric Inhibitor of the Uridine Diphosphate N‑Acetylglucosamine Pyrophosphorylase from Trypanosoma brucei PY - 2015/12/16 AU - Michael D. Urbaniak AU - Iain T. Collie AU - Wenxia Fang AU - Tonia Aristotelous AU - Susanne Eskilsson AU - Olawale G. Raimi AU - Justin Harrison AU - Iva Hopkins Navratilova AU - Julie A. Frearson AU - Daan M. F. van Aalten AU - Michael A. J. Ferguson UR - https://acs.figshare.com/articles/journal_contribution/A_Novel_Allosteric_Inhibitor_of_the_Uridine_Diphosphate_i_N_i_Acetylglucosamine_Pyrophosphorylase_from_i_Trypanosoma_brucei_i_/2025816 DO - 10.1021/cb400411x.s001 L4 - https://ndownloader.figshare.com/files/3597108 KW - bind UTP KW - Structural characterization KW - Trypanosoma brucei KW - parasite enzyme KW - brucei UAP KW - Novel Allosteric Inhibitor KW - etiological agent KW - allosteric binding site KW - site residues KW - Biophysical characterization KW - trypanosome enzymes KW - substrate binding KW - UAP enzyme kinetics KW - inhibitor KW - allosteric site N2 - 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. ER -