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Comprehensive analysis of flagellin glycosylation in Campylobacter jejuni NCTC 11168 reveals incorporation of legionaminic acid and its importance for host colonization

Zebian, Najwa; Merkx-Jacques, Alexandra; Pittock, Paula P.; Houle, Sébastien; Dozois, Charles M; Lajoie, Gilles A; Creuzenet, Carole (2016). Comprehensive analysis of flagellin glycosylation in Campylobacter jejuni NCTC 11168 reveals incorporation of legionaminic acid and its importance for host colonization Glycobiology , vol. 26 , nº 4. p. 386-397. DOI: 10.1093/glycob/cwv104.

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Résumé

Campylobacter jejuni is the leading cause of bacterial gastroenteritis. It relies on several virulence factors for host colonization, including glycosylated flagella. C. jejuni NCTC 11168 modifies its flagellins with pseudaminic acid derivatives. It is also presumed to modify these proteins with legionaminic acid, although no glycopeptide evidence was available at the onset of this study. The enzyme encoded by cj1319 can be used to make legionaminic acid in vitro, but the pathway for legionaminic acid synthesis partially inferred by knockout mutagenesis in Campylobacter coli VC167 excludes Cj1319. To address this contradiction, we examined the presence of legionaminic acid in flagellin glycopeptides of wild type C. jejuni NCTC 11168 and of a cj1319 knockout mutant. We used high-energy collision induced dissociation to obtain amino acid sequences while also visualizing signature sugar oxonium ions. Data analysis was performed with PEAKS software and spectra were manually inspected for glycopeptide determination and verification. We showed that legionaminic acid is present on the flagellins of C. jejuni NCTC 11168 and that flagellin glycosylation is highly heterogeneous, with up to 6 different sugars singly present at a given site. We found that the cj1319 mutant produces more legionaminic acid than wild type, thus excluding the requirement for Cj1319 for legionaminic acid synthesis. We also showed that this mutant has enhanced chicken colonization compared to wild type, which may in part be attributed to the high content of legionaminic acid on its flagella.

Type de document:
Mots-clés libres: Campylobacter jejuni; flagellins; glycopeptide; legionaminic acid; pseudaminic acid
Centre: Centre INRS-Institut Armand Frappier
Date de dépôt: 16 mai 2017 18:54
Dernière modification: 16 mai 2017 18:54
URI: http://espace.inrs.ca/id/eprint/3278

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