Structural and Functional Characterization of the Clostridium perfringens N-Acetylmannosamine-6-phosphate 2-Epimerase Essential for the Sialic Acid Salvage Pathway
In: Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (51), pp.35215-35224. ⟨10.1074/jbc.m114.604272⟩ Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (51), pp.35215--24. ⟨10.1074/jbc.M114.604272⟩ Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (51), pp.35215-35224. ⟨10.1074/jbc.M114.604272⟩ Journal of Biological Chemistry, 2014, 289 (51), pp.35215-35224. ⟨10.1074/jbc.m114.604272⟩; (2014)
Online
unknown
Zugriff:
International audience; Pathogenic bacteria are endowed with an arsenal of specialized enzymes to convert nutrient compounds from their cell hosts. The essential N-acetylmannosamine-6-phosphate 2-epimerase (NanE) belongs to a convergent glycolytic pathway for utilization of the three amino sugars, GlcNAc, ManNAc, and sialic acid. The crystal structure of ligand-free NanE from Clostridium perfringens reveals a modified triose-phosphate isomerase (?/?)8 barrel in which a stable dimer is formed by exchanging the C-terminal helix. By retaining catalytic activity in the crystalline state, the structure of the enzyme bound to the GlcNAc-6P product identifies the topology of the active site pocket and points to invariant residues Lys(66) as a putative single catalyst, supported by the structure of the catalytically inactive K66A mutant in complex with substrate ManNAc-6P. (1)H NMR-based time course assays of native NanE and mutated variants demonstrate the essential role of Lys(66) for the epimerization reaction with participation of neighboring Arg(43), Asp(126), and Glu(180) residues. These findings unveil a one-base catalytic mechanism of C2 deprotonation/reprotonation via an enolate intermediate and provide the structural basis for the development of new antimicrobial agents against this family of bacterial 2-epimerases.
Titel: |
Structural and Functional Characterization of the Clostridium perfringens N-Acetylmannosamine-6-phosphate 2-Epimerase Essential for the Sialic Acid Salvage Pathway
|
---|---|
Autor/in / Beteiligte Person: | Bourne, Yves ; Vincent, Florence ; Brannigan, James A. ; Sebban-Kreuzer, Corinne ; Pelissier, Marie-Cécile ; Guerlesquin, Françoise ; Architecture et fonction des macromolécules biologiques (AFMB) ; Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Institut National de la Recherche Agronomique (INRA) ; Laboratoire d'ingénierie des systèmes macromoléculaires (LISM) ; Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU) ; University of York [York, UK] ; ANR-10-INTB-1501,BioPol folders,Repliement et stabilisation par des polymeres amphiphiles biocompatibles de fragments scFv marqueurs cellulaires de cancers.(2010) ; UNIROUEN - UFR Santé (UNIROUEN UFR Santé) ; Université de Rouen Normandie (UNIROUEN) ; Normandie Université (NU)-Normandie Université (NU) ; Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS) ; Architecture et fonction des Macromolécules Biologiques - UMR 6098 (AFMB) ; Université de Provence - Aix-Marseille 1-Centre National de la Recherche Scientifique (CNRS) ; Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE) ; Institut National de la Recherche Agronomique (INRA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS) ; Programme Blanc International édition 2010 - Repliement et stabilisation par des polymeres amphiphiles biocompatibles de fragments scFv marqueurs cellulaires de cancers. - - BioPol folders2010 - ANR-10-INTB-1501 - Blanc international 2010 -, VALID |
Link: | |
Quelle: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (51), pp.35215-35224. ⟨10.1074/jbc.m114.604272⟩ Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (51), pp.35215--24. ⟨10.1074/jbc.M114.604272⟩ Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2014, 289 (51), pp.35215-35224. ⟨10.1074/jbc.M114.604272⟩ Journal of Biological Chemistry, 2014, 289 (51), pp.35215-35224. ⟨10.1074/jbc.m114.604272⟩; (2014) |
Veröffentlichung: | HAL CCSD, 2014 |
Medientyp: | unknown |
ISSN: | 0021-9258 (print) ; 1083-351X (print) |
Schlagwort: |
|
Sonstiges: |
|