Simultaneous Glycan-peptide Characterization Using Hydrophilic Interaction Chromatography And Parallel Fragmentation By Cid, Higher Energy Collisional Dissociation, And Electron Transfer Dissociation Ms Applied To The N-linked Glycoproteome Of Campyl

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dc.contributor.author Scott, Nichollas en_US
dc.contributor.author Parker, B.L. en_US
dc.contributor.author Connolly, A.M. en_US
dc.contributor.author Paulech, J. en_US
dc.contributor.author Edwards, Alistair en_US
dc.contributor.author Crossett, B. en_US
dc.contributor.author Falconer, Delia en_US
dc.contributor.author Kolarich, D. en_US
dc.contributor.author Djordjevic, Steven en_US
dc.contributor.author Hojrup, P. en_US
dc.contributor.author Packer, N.H. en_US
dc.contributor.author Larsen, Martin en_US
dc.contributor.author Cordwell, Stuart en_US
dc.contributor.editor en_US
dc.date.accessioned 2012-10-12T03:34:30Z
dc.date.available 2012-10-12T03:34:30Z
dc.date.issued 2011 en_US
dc.identifier 2011003485 en_US
dc.identifier.citation Scott N.E. et al. 2011, 'Simultaneous Glycan-peptide Characterization Using Hydrophilic Interaction Chromatography And Parallel Fragmentation By Cid, Higher Energy Collisional Dissociation, And Electron Transfer Dissociation Ms Applied To The N-linked Glycoproteome Of Campyl', American Society for Biochemistry and Molecular Biology, Inc., vol. 10, no. 2, pp. M000031 en_US
dc.identifier.issn 1535-9476 en_US
dc.identifier.other C1 en_US
dc.identifier.uri http://hdl.handle.net/10453/18680
dc.description.abstract Campylobacter jejuni is a gastrointestinal pathogen that is able to modify membrane and periplasmic proteins by the N-linked addition of a 7-residue glycan at the strict attachment motif (D/E) XNX(S/T). Strategies for a comprehensive analysis of the targets of glycosylation, however, are hampered by the resistance of the glycan-peptide bond to enzymatic digestion or beta-elimination and have previously concentrated on soluble glycoproteins compatible with lectin affinity and gel-based approaches. We developed strategies for enriching C. jejuni HB93-13 glycopeptides using zwitterionic hydrophilic interaction chromatography and examined novel fragmentation, including collision-induced dissociation ( CID) and higher energy collisional (C-trap) dissociation (HCD) as well as CID/electron transfer dissociation (ETD) mass spectrometry. CID/HCD enabled the identification of glycan structure and peptide backbone, allowing glycopeptide identification, whereas CID/ETD enabled the elucidation of glycosylation sites by maintaining the glycan-peptide linkage. A total of 130 glycopeptides, representing 75 glycosylation sites, were identified from LC-MS/MS using zwitterionic hydrophilic interaction chromatography coupled to CID/HCD and CID/ETD. CID/HCD provided the majority of the identifications (73 sites) compared with ETD (26 sites). We also examined soluble glycoproteins by soybean agglutinin affinity and two-dimensional electrophoresis and identified a further six glycosylation sites. This study more than doubles the number of confirmed N-linked glycosylation sites in C. jejuni and is the first to utilize HCD fragmentation for glycopeptide identification with intact glycan. We also show that hydrophobic integral membrane proteins are significant targets of glycosylation in this organism. en_US
dc.language en_US
dc.publisher American Society for Biochemistry and Molecular Biology, Inc. en_US
dc.title Simultaneous Glycan-peptide Characterization Using Hydrophilic Interaction Chromatography And Parallel Fragmentation By Cid, Higher Energy Collisional Dissociation, And Electron Transfer Dissociation Ms Applied To The N-linked Glycoproteome Of Campyl en_US
dc.parent Molecular And Cellular Proteomics en_US
dc.journal.volume 10 en_US
dc.journal.number 2 en_US
dc.publocation USA en_US
dc.identifier.startpage 1 en_US
dc.identifier.endpage en_US
dc.identifier.endpage 12 en_US
dc.cauo.name SCI.Gore Hill Research Laboratory en_US
dc.conference Verified OK en_US
dc.for 110899 en_US
dc.personcode 0000052321 en_US
dc.personcode 0000075469 en_US
dc.personcode 0000075470 en_US
dc.personcode 0000075471 en_US
dc.personcode 0000063289 en_US
dc.personcode 0000075473 en_US
dc.personcode 034107 en_US
dc.personcode 0000075475 en_US
dc.personcode 107126 en_US
dc.personcode 0000075477 en_US
dc.personcode 0000075478 en_US
dc.personcode 0000063293 en_US
dc.personcode 0000027041 en_US
dc.percentage 100 en_US
dc.classification.name Medical Microbiology not elsewhere classified en_US
dc.classification.type FOR-08 en_US
dc.edition en_US
dc.custom en_US
dc.date.activity en_US
dc.location.activity en_US
dc.description.keywords HUMAN EPITHELIAL-CELLS; MASS-SPECTROMETRY; PROTEIN GLYCOSYLATION; POSTTRANSLATIONAL MODIFICATIONS; LIQUID-CHROMATOGRAPHY; CONSENSUS SEQUENCE; IDENTIFICATION; COLONIZATION; EXPRESSION; PROTEOMICS en_US
dc.staffid en_US


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