title: Characterization of the structurally diverse N-linked glycans of Campylobacter species.
abstract: The Gram-negative bacterium Campylobacter jejuni encodes an extensively characterized N-linked protein glycosylation system that modifies many surface proteins with a heptasaccharide glycan. In C. jejuni, the genes that encode the enzymes required for glycan biosynthesis and transfer to protein are located at a single pgl gene locus. Similar loci are also present in the genome sequences of all other Campylobacter species, although variations in gene content and organization are evident. In this study, we have demonstrated that only Campylobacter species closely related to C. jejuni produce glycoproteins that interact with both a C. jejuni N-linked-glycan-specific antiserum and a lectin known to bind to the C. jejuni N-linked glycan. In order to further investigate the structure of Campylobacter N-linked glycans, we employed an in vitro peptide glycosylation assay combined with mass spectrometry to demonstrate that Campylobacter species produce a range of structurally distinct N-linked glycans with variations in the number of sugar residues (penta-, hexa-, and heptasaccharides), the presence of branching sugars, and monosaccharide content. These data considerably expand our knowledge of bacterial N-linked glycan structure and provide a framework for investigating the role of glycosyltransferases and sugar biosynthesis enzymes in glycoprotein biosynthesis with practical implications for synthetic biology and glycoengineering.
authors.affiliation: Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.
authors.forename: Adrian J
authors.initials: AJ
authors.lastname: Jervis
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Butler
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Lawson
authors.forename: Rebecca
authors.initials: R
authors.lastname: Langdon
authors.forename: Brendan W
authors.initials: BW
authors.lastname: Wren
authors.forename: Dennis
authors.initials: D
authors.lastname: Linton
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002167:Campylobacter / Q000235:genetics / Q000378:metabolism*; D002236:Carbohydrate Conformation; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D014644:Genetic Variation; D006031:Glycosylation; D010802:Phylogeny; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism*; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001426:Bacterial Proteins; D011134:Polysaccharides