Identification of the carbohydrate moieties and glycosylation motifs in Campylobacter jejuni flagellin: J.Biol.Chem.

Download
  1. Get@NRC: Identification of the carbohydrate moieties and glycosylation motifs in Campylobacter jejuni flagellin: J.Biol.Chem. (Opens in a new window)
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Journal titleJ.Biol.Chem.
Volume276
Issue37
Pages3486234870; # of pages: 9
SubjectACID; Amino Acid Sequence; analysis; biosynthesis; Campylobacter; Campylobacter coli; Campylobacter jejuni; Canada; capillary; carbohydrate; chemistry; Chromatography; DNA; DOMAIN; Flagellin; GENE; glycopeptide; Glycopeptides; Glycosylation; HETEROGENEITY; IDENTIFICATION; mass spectrometry; MASS-SPECTROMETRY; MOLECULAR; Molecular Sequence Data; peptide; Peptides; protein; RESIDUES; RESTRICTED; SEQUENCE; SITE; SPECTROSCOPY; Spectrum Analysis,Mass; STRAIN; STRAINS; Support,U.S.Gov't,P.H.S.; surface
AbstractFlagellins from three strains of Campylobacter jejuni and one strain of Campylobacter coli were shown to be extensively modified by glycosyl residues, imparting an approximate 6000-Da shift from the molecular mass of the protein predicted from the DNA sequence. Tryptic peptides from C. jejuni 81-176 flagellin were subjected to capillary liquid chromatography-electrospray mass spectrometry with a high/low orifice stepping to identify peptide segments of aberrant masses together with their corresponding glycosyl appendages. These modified peptides were further characterized by tandem mass spectrometry and preparative high performance liquid chromatography followed by nano-NMR spectroscopy to identify the nature and precise site of glycosylation. These analyses have shown that there are 19 modified Ser/Thr residues in C. jejuni 81-176 flagellin. The predominant modification found on C. jejuni flagellin was O-linked 5,7-diacetamido-3,5,7,9-tetradeoxy-l-glycero-l-manno-nonulosonic acid (pseudaminic acid, Pse5Ac7Ac) with additional heterogeneity conferred by substitution of the acetamido groups with acetamidino and hydroxyproprionyl groups. In C. jejuni 81-176, the gene Cj1316c, encoding a protein of unknown function, was shown to be involved in the biosynthesis and/or the addition of the acetamidino group on Pse5Ac7Ac. Glycosylation is not random, since 19 of the total 107 Ser/Thr residues are modified, and all but one of these are restricted to the central, surface-exposed domain of flagellin when folded in the filament. The mechanism of attachment appears unrelated to a consensus peptide sequence but is rather based on surface accessibility of Ser/Thr residues in the folded protein
Publication date
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedNo
NRC numberTHIBAULT2001
NPARC number9371780
Export citationExport as RIS
Report a correctionReport a correction
Record identifierc8679f63-99f9-4516-9cca-71df93ef4f03
Record created2009-07-10
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)