title: Liquid chromatography-tandem mass spectrometry-based fragmentation analysis of glycopeptides.
abstract: The use of liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS(n)) for the glycoproteomic characterization of glycopeptides is a growing field of research. The N- and O-glycosylated peptides (N- and O-glycopeptides) analyzed typically originate from protease-digested glycoproteins where many of them are expected to be biomedically important. Examples of LC-MS(2) and MS(3) fragmentation strategies used to pursue glycan structure, peptide identity and attachment-site identification analyses of glycopeptides are described in this review. MS(2) spectra, using the CID and HCD fragmentation techniques of a complex biantennary N-glycopeptide and a core 1 O-glycopeptide, representing two examples of commonly studied glycopeptide types, are presented. A few practical tips for accomplishing glycopeptide analysis using reversed-phase LC-MS(n) shotgun proteomics settings, together with references to the latest glycoproteomic studies, are presented.
authors.affiliation: Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska Academy at the University of Gothenburg, Gothenburg, Sweden. jonas.nilsson@clinchem.gu.se.
authors.forename: Jonas
authors.initials: J
authors.lastname: Nilsson
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods*; D006020:Glycopeptides / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D006020:Glycopeptides
keywords: Collision-induced dissociation; Electron capture dissociation; Electron transfer dissociation; Glycopeptides; Glycoproteomics; Higher-energy collisional dissociation; Mass spectrometry; Oxonium ions