title: Characterization of protein N-glycosylation by tandem mass spectrometry using complementary fragmentation techniques.
abstract: The analysis of post-translational modifications (PTMs) by proteomics is regarded as a technically challenging undertaking. While in recent years approaches to examine and quantify protein phosphorylation have greatly improved, the analysis of many protein modifications, such as glycosylation, are still regarded as problematic. Limitations in the standard proteomics workflow, such as use of suboptimal peptide fragmentation methods, can significantly prevent the identification of glycopeptides. The current generation of tandem mass spectrometers has made available a variety of fragmentation options, many of which are becoming standard features on these instruments. We have used three common fragmentation techniques, namely CID, HCD, and ETD, to analyze a glycopeptide and highlight how an integrated fragmentation approach can be used to identify the modified residue and characterize the N-glycan on a peptide.
authors.affiliation: ARC Centre of Excellence in Plant Cell Walls, School of BioSciences, The University of Melbourne Melbourne, VIC, Australia.
authors.forename: Kristina L
authors.initials: KL
authors.lastname: Ford
authors.affiliation: ARC Centre of Excellence in Plant Cell Walls, School of BioSciences, The University of Melbourne Melbourne, VIC, Australia.
authors.forename: Wei
authors.initials: W
authors.lastname: Zeng
authors.affiliation: ARC Centre of Excellence in Plant Cell Walls, School of BioSciences, The University of Melbourne Melbourne, VIC, Australia ; Physical Biosciences Division, Joint BioEnergy Institute, Lawrence Berkeley National Laboratory Berkeley, CA, USA.
authors.forename: Joshua L
authors.initials: JL
authors.lastname: Heazlewood
authors.affiliation: ARC Centre of Excellence in Plant Cell Walls, School of BioSciences, The University of Melbourne Melbourne, VIC, Australia.
authors.forename: Antony
authors.initials: A
authors.lastname: Bacic
keywords: electron-transfer dissociation; fragmentation; glycosylation; post-translational modification; tandem mass spectrometry