title: Determination of the MurD mechanism through crystallographic analysis of enzyme complexes.
abstract: UDP -N- acetylmuramoyl- L -alanine: D -glutamate (MurD) ligase catalyses the addition of d -glutamate to the nucleotide precursor UDP -N- acetylmuramoyl- L -alanine (UMA). The crystal structures of three complexes of Escherichia coli MurD with a variety of substrates and products have been determined to high resolution. These include (1) the quaternary complex of MurD, the substrate UMA, the product ADP, and Mg2+, (2) the quaternary complex of MurD, the substrate UMA, the product ADP, and Mn2+, and (3) the binary complex of MurD with the product UDP - N- acetylmuramoyl- L -alanine- D -glutamate (UMAG). The reaction mechanism supported by these structures proceeds by the phosphorylation of the C-terminal carboxylate group of UMA by the gamma-phosphate group of ATP to form an acyl-phosphate intermediate, followed by the nucleophilic attack by the amino group of D-glutamate to produce UMAG. A key feature in the reaction intermediate is the presence of two magnesium ions bridging negatively charged groups.
authors.affiliation: Laboratoire de Cristallographie Macromoléculaire, Institut de Biologie Structurale Jean-Pierre Ebel (CEA-CNRS), 41 rue Jules Horowitz, Grenoble Cedex 1, F-38027, France.
authors.forename: J A
authors.initials: JA
authors.lastname: Bertrand
authors.forename: G
authors.initials: G
authors.lastname: Auger
authors.forename: L
authors.initials: L
authors.lastname: Martin
authors.forename: E
authors.initials: E
authors.lastname: Fanchon
authors.forename: D
authors.initials: D
authors.lastname: Blanot
authors.forename: D
authors.initials: D
authors.lastname: Le Beller
authors.forename: J
authors.initials: J
authors.lastname: van Heijenoort
authors.forename: O
authors.initials: O
authors.lastname: Dideberg
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D018360:Crystallography, X-Ray; D004151:Dipeptides / Q000737:chemistry / Q000378:metabolism; D006868:Hydrolysis; D008274:Magnesium / Q000737:chemistry / Q000378:metabolism; D008345:Manganese / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010453:Peptide Synthases / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism
chemical_list: D004151:Dipeptides; C120188:UDP-N-acetylmuramoylalanyl-glutamate; D014538:Uridine Diphosphate N-Acetylmuramic Acid; D000266:Adenylyl Imidodiphosphate; D008345:Manganese; D000244:Adenosine Diphosphate; D010453:Peptide Synthases; C053379:UDP-N-acetylmuramoylalanine-D-glutamate ligase; D008274:Magnesium