Structure of the N -linked glycan present on multiple glycoproteins in the gram-negative bacterium, Campylobacter jejuni: J.Biol.Chem.

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TypeArticle
Journal titleJournal Of Biological Chemistry
Volume277
Issue45
Pages4253042539; # of pages: 10
Subjectanalysis; Bacteria; biosynthesis; Campylobacter; Campylobacter jejuni; Chromatography; Digestion; ENZYME; GENE; Genes; Glycine; glycopeptide; Glycopeptides; glycoprotein; Glycoproteins; Glycosylation; Gram Negative Bacteria; Gram-Negative Bacteria; HIV; IDENTIFICATION; lectin; LED; mass spectrometry; MASS-SPECTROMETRY; Mixture; Multiple; N-linked; Neisseria; Neisseria meningitidis; Pronase; protein; PROTEIN GLYCOSYLATION; Proteins; structure; technique
AbstractMass spectrometry investigations of partially purified C. jejuni protein PEB3 showed it to be partially modified with an Asn-linked glycan of mass 1406 Da and comprised of one hexose, five HexNAc and a species of mass 228 Da, consistent with a trideoxydiacetamidohexose. By means of soybean lectin affinity chromatography, a mixture of glycoproteins was obtained from a glycine extract, and 2D-gel proteomics analysis led to the identification of at least 22 glycoproteins, predominantly annotated as periplasmic proteins. Glycopeptides were prepared from the glycoprotein mixture by pronase digestion and gel-filtration. The structure of the glycan was determined by using nano-NMR techniques to be GalNAc-a1,4-GalNAc-a1,4-GalNAc-a1,4-[Glcb1,3-]GalNAc-a1,4-GalNAc-a1,3-Bac- b1,N-Asn-Xaa, where Bac is bacillosamine, 2,4-diacetamido-2,4,6-trideoxyglucopyranose. Protein glycosylation was abolished when the pglB gene was mutated, providing further evidence that the enzyme encoded by this gene is responsible for formation of the glycopeptide N-linkage. Comparison of the pgl locus with that of Neisseria meningitidis suggested that most of the homologous genes are probably involved in the biosynthesis of bacillosamine
Publication date
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedNo
NRC numberYOUNG2002
NPARC number9383570
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Record identifier05300de1-6c08-4f44-9602-1df163aaa4f5
Record created2009-07-10
Record modified2016-05-09
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