title: Characterizing Glycoproteins by Mass Spectrometry in Campylobacter jejuni.
abstract: The glycosylation systems of Campylobacter jejuni (C. jejuni) are considered archetypal examples of both N- and O-linked glycosylations in the field of bacterial glycosylation. The discovery and characterization of these systems both have revealed important biological insight into C. jejuni and have led to the refinement and enhancement of methodologies to characterize bacterial glycosylation. In general, mass spectrometry-based characterization has become the preferred methodology for the study of C. jejuni glycosylation because of its speed, sensitivity, and ability to enable both qualitative and quantitative assessments of glycosylation events. In these experiments the generation of insightful data requires the careful selection of experimental approaches and mass spectrometry (MS) instrumentation. As such, it is essential to have a deep understanding of the technologies and approaches used for characterization of glycosylation events. Here we describe protocols for the initial characterization of C. jejuni glycoproteins using protein-/peptide-centric approaches and discuss considerations that can enhance the generation of insightful data.
authors.affiliation: Department of Microbiology and Immunology, Doherty Institute, The University of Melbourne, 792 Elizabeth St., Melbourne, Victoria, 3001, Australia. nichollas.scott@unimelb.edu.au.
authors.forename: Nichollas E
authors.initials: NE
authors.lastname: Scott
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000302:isolation & purification* / Q000378:metabolism; D016123:Campylobacter jejuni / Q000737:chemistry* / Q000254:growth & development / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000302:isolation & purification* / Q000378:metabolism; D006031:Glycosylation; D013058:Mass Spectrometry / Q000379:methods; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods*; D059748:Proteolysis; D014357:Trypsin / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D006023:Glycoproteins; D010446:Peptide Fragments; D014357:Trypsin
keywords: Campylobacter jejuni; Glycoprotein; Mass spectrometry