title: Direct determination of glycosylation sites in O-fucosylated glycopeptides using nano-electrospray quadrupole time-of-flight mass spectrometry.
abstract: O-Fucosylation is an unusual posttranslational modification present in several proteins that play important roles in physiological processes such as coagulation, cell signaling and metastasis. Although the exact function of the modification is still unclear, the number of proteins found to be modified is increasing, and there is a need for further structural and functional analyses. Here we report on a rapid and straightforward approach in the analysis of glycosylation status and determination of glycosylation sites in O-fucosylated glycopeptides using nano-electrospray quadrupole time-of-flight (nano-ESI Q-TOF) mass spectrometry. In a single measurement of previously chemically untreated O-fucosylated peptides originating from the thrombospondin-1 repeats, we were able to determine the glycosylation status of the analyzed peptide, the glycosylation site, and the glycan structure. The abundance of glycosylated peptide fragment ions in MS(2) spectra suggests that nano-ESI Q-TOF mass spectrometry can be used as a general approach in structural studies of O-fucosylation in proteins.
authors.affiliation: Institute for Medical Physics and Biophysics, Department for Biomedical Analysis, University of Münster, Robert-Koch-Str. 31, D-48149 Münster, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Macek
authors.forename: J
authors.initials: J
authors.lastname: Hofsteenge
authors.forename: J
authors.initials: J
authors.lastname: Peter-Katalinić
mesh_terms: D000818:Animals; D006020:Glycopeptides / Q000737:chemistry*; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D019700:Thrombospondin 1 / Q000737:chemistry
chemical_list: D006020:Glycopeptides; D011993:Recombinant Fusion Proteins; D019700:Thrombospondin 1