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Targeted metabolomics analysis of Campylobacter coli VC167 reveals legionaminic acid derivatives as novel flagellar glycans

Par Conseil national de recherches du Canada

DOITrouver le DOI : https://doi.org/10.1074/jbc.M611027200
AuteurRechercher : 1; Rechercher : 1; Rechercher : 2; Rechercher : 1; Rechercher : 1; Rechercher : ; Rechercher : ; Rechercher : 1; Rechercher : 1; Rechercher : 2
Affiliation
  1. Conseil national de recherches Canada. Institut des sciences biologiques du CNRC
  2. Conseil national de recherches Canada. Institut des biosciences marines du CNRC
FormatTexte, Article
Sujetbiosynthetic pathways; Campylobacter coli; Campylobacter jejuni; Canada; chemistry; chromatography, liquid; complement; cyclic amp; enzymes; flagellin; gene expression regulation, bacterial; genes, bacterial; genetics; glycans; glycosylation; interaction; ions; magnetic resonance spectroscopy; mass spectrometry; metabolism; models, chemical; models, molecular; molecular sequence data; mutagenesis; mutants; mutation; NMR spectroscopy; nucleotides; pathway; polysaccharides; protein; role; sialic acids; spectrometry, mass, matrix-assisted laser desorption-ionization; spectroscopy; structural analysis; sugars
Résumé
Glycosylation of Campylobacter flagellin is required for the biogenesis of a functional flagella filament. Recently, we used a targeted metabolomics approach using mass spectrometry and NMR to identify changes in the metabolic profile of wild type and mutants in the flagellar glycosylation locus, characterize novel metabolites, and assign function to genes to define the pseudaminic acid biosynthetic pathway in Campylobacter jejuni 81-176 (McNally, D. J., Hui, J. P., Aubry, A. J., Mui, K. K., Guerry, P., Brisson, J. R., Logan, S. M., and Soo, E. C. (2006) J. Biol. Chem. 281, 18489-18498). In this study, we use a similar approach to further define the glycome and metabolomic complement of nucleotide-activated sugars in Campylobacter coli VC167. Herein we demonstrate that, in addition to CMP-pseudaminic acid, C. coli VC167 also produces two structurally distinct nucleotide-activated nonulosonate sugars that were observed as negative ions at m/z 637 and m/z 651 (CMP-315 and CMP-329). Hydrophilic interaction liquid chromatography-mass spectrometry yielded suitable amounts of the pure sugar nucleotides for NMR spectroscopy using a cold probe. Structural analysis in conjunction with molecular modeling identified the sugar moieties as acetamidino and N-methylacetimidoyl derivatives of legionaminic acid (Leg5Am7Ac and Leg5AmNMe7Ac). Targeted metabolomic analyses of isogenic mutants established a role for the ptmA-F genes and defined two new ptm genes in this locus as legionaminic acid biosynthetic enzymes. This is the first report of legionaminic acid in Campylobacter sp. and the first report of legionaminic acid derivatives as modifications on a protein
Date de publication
Maison d’éditionAmerican Society for Biochemistry and Molecular Biology
Dans
  • The Journal of Biological Chemistry 282, nº 19 : 14463–14475.
Langueanglais
Publications évaluées par des pairsOui
Numéro du CNRCMCNALLY2007A
1702
Numéro NPARC9362866
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Identificateur de l’enregistrementbc8fa29a-3a30-4ae1-94ef-2153a3dd23bb
Enregistrement créé2009-07-10
Enregistrement modifié2020-05-10
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