title: Purification and biochemical characterisation of GlmU from Yersinia pestis.
abstract: Antibiotic resistance has emerged as a real threat to mankind, rendering many compounds ineffective in the fight against bacterial infection, including for significant diseases such as plague caused by Yersinia pestis. Essential genes have been identified as promising targets for inhibiting with new classes of compounds. Previously, the gene encoding the bifunctional UDP-N-acetylglucosamine pyrophosphorylase/glucosamine-1-phosphate N-acetyltransferase enzyme GlmU was confirmed as an essential gene in Yersinia. As a step towards exploiting this target for antimicrobial screening, we undertook a biochemical characterisation of the Yersinia GlmU. Effects of pH and magnesium concentration on the acetyltransferase and uridyltransferase activities were analysed, and kinetic parameters were determined. The acetyltransferase activity, which is strongly increased in the presence of reducing agent, was shown to be susceptible to oxidation and thiol-specific reagents.
authors.affiliation: Laboratoire des Enveloppes Bactériennes et Antibiotiques, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, UMR 8619 CNRS, Bâtiment 430, Université Paris-Sud, 91405, Orsay, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Patin
authors.forename: Marc
authors.initials: M
authors.lastname: Bayliss
authors.forename: Dominique
authors.initials: D
authors.lastname: Mengin-Lecreulx
authors.forename: Petra
authors.initials: P
authors.lastname: Oyston
authors.forename: Didier
authors.initials: D
authors.lastname: Blanot
mesh_terms: D000123:Acetyltransferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D000595:Amino Acid Sequence; D004789:Enzyme Activation / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology; D008969:Molecular Sequence Data; D009713:Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D016415:Sequence Alignment; D015010:Yersinia pestis / Q000201:enzymology* / Q000235:genetics
chemical_list: D016877:Oxidants; D008623:Mercaptoethanol; D000123:Acetyltransferases; C428383:glucosamine-1-phosphate acetyltransferase; D009713:Nucleotidyltransferases; C023226:UDPacetylglucosamine pyrophosphorylase; D008274:Magnesium