title: A comparison of energy-resolved vibrational activation/dissociation characteristics of protonated and sodiated high mannose N-glycopeptides.
abstract: Fragmentation of glycopeptides in tandem mass spectrometry (MS/MS) plays a pivotal role in site-specific protein glycosylation profiling by allowing specific oligosaccharide compositions and connectivities to be associated with specific loci on the corresponding protein. Although MS/MS analysis of glycopeptides has been successfully performed using a number of distinct ion dissociation methods, relatively little is known regarding the fragmentation characteristics of glycopeptide ions with various charge carriers. In this study, energy-resolved vibrational activation/dissociation was examined via collision-induced dissociation for a group of related high mannose tryptic glycopeptides as their doubly protonated, doubly sodiated, and hybrid protonated sodium adduct ions. The doubly protonated glycopeptide ions with various compositions were found to undergo fragmentation over a relatively low but wide range of collision energies compared with the doubly sodiated and hybrid charged ions, and were found to yield both glycan and peptide fragmentation depending on the applied collision energy. By contrast, the various doubly sodiated glycopeptides were found to dissociate over a significantly higher but narrow range of collision energies, and exhibited only glycan cleavages. Interestingly, the hybrid protonated sodium adduct ions were consistently the most stable of the precursor ions studied, and provided fragmentation information spanning both the glycan and the peptide moieties. Taken together, these findings illustrate the influence of charge carrier over the energy-resolved vibrational activation/dissociation characteristics of glycopeptides, and serve to suggest potential strategies that exploit the analytically useful features uniquely afforded by specific charge carriers or combinations thereof.
authors.affiliation: Department of Chemistry, University of Nebraska-Lincoln, 711 Hamilton Hall, Lincoln, NE, 68588-0304, USA.
authors.forename: Forouzan
authors.initials: F
authors.lastname: Aboufazeli
authors.forename: Venkata
authors.initials: V
authors.lastname: Kolli
authors.forename: Eric D
authors.initials: ED
authors.lastname: Dodds
mesh_terms: D006020:Glycopeptides / Q000737:chemistry*; D007477:Ions / Q000737:chemistry; D008358:Mannose / Q000737:chemistry*; D011522:Protons; D012964:Sodium / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014732:Vibration
chemical_list: D006020:Glycopeptides; D007477:Ions; D011522:Protons; D012964:Sodium; D008358:Mannose