Direct determination of glycosylation sites in O-fucosylated glycopeptides using nano-electrospray quadrupole time-of-flight mass spectrometry

Rapid Communications in Mass Spectrometry : RCM
B MacekJ Peter-Katalinic

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.

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Citations

Jul 22, 2008·Glycoconjugate Journal·François FenailleNicolas Bihoreau
Nov 3, 2004·Journal of the American Society for Mass Spectrometry·Alina ZamfirMark Allen
Feb 14, 2002·Chemical Reviews·Yehia Mechref, Milos V Novotny
Aug 26, 2010·Molecular & Cellular Proteomics : MCP·Sheng PanTeresa A Brentnall
Aug 4, 2004·Journal of Bioinformatics and Computational Biology·P Kearney, P Thibault
Nov 16, 2011·International Journal of Proteomics·Evelyn H Kim, David E Misek
Sep 25, 2004·Mass Spectrometry Reviews·Helen J CooperAlan G Marshall
Oct 20, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Manfred WuhrerCornelis H Hokke
Nov 23, 2006·Biochimica Et Biophysica Acta·Bogdan A BudnikJudith A J Steen
Sep 30, 2003·Rapid Communications in Mass Spectrometry : RCM·Michael Mormann, Jasna Peter-Katalinić
Feb 18, 2004·Mass Spectrometry Reviews·Joseph Zaia
Jan 18, 2006·Methods in Enzymology·Jasna Peter-Katalinić
Nov 30, 2006·Methods in Enzymology·Aleksandra Nita-Lazar, Robert S Haltiwanger
Oct 13, 2001·Journal of Mass Spectrometry : JMS
Dec 3, 2005·European Journal of Mass Spectrometry·Michael MormannJasna Peter-Katalinić
Jul 4, 2002·Molecular & Cellular Proteomics : MCP·Anne Gonzalez de PeredoJan Hofsteenge
Aug 2, 2017·PLoS Neglected Tropical Diseases·Kristian E SwearingenSebastian A Mikolajczak
Jul 4, 2009·Journal of Proteome Research·François FenailleEric Ezan
Apr 1, 2004·Analytical Chemistry·Alina ZamfirJasna Peter-Katalinić

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