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Analysis of protein glycosylation by mass spectrometry

Analysis of protein glycosylation by mass spectrometry We present a detailed protocol for the structural analysis of protein-linked glycans. In this approach, appropriate for glycomics studies, N-linked glycans are released using peptide N-glycosidase F and O-linked glycans are released by reductive alkaline β-elimination. Using strategies based on mass spectrometry (matrix-assisted laser desorption/ionization–time of flight mass spectrometry and nano-electrospray ionization mass spectrometry/mass spectrometry (nano-ESI-MS-MS)), chemical derivatization, sequential exoglycosidase digestions and linkage analysis, the structures of the N- and/or O-glycans are defined. This approach can be used to study the glycosylation of isolated complex glycoproteins or of numerous glycoproteins encountered in a complex biological medium (cells, tissues and physiological fluids). http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Protocols Springer Journals

Analysis of protein glycosylation by mass spectrometry

Nature Protocols , Volume 2 (7) – Jun 21, 2007

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References (47)

Publisher
Springer Journals
Copyright
Copyright © 2007 by Nature Publishing Group
Subject
Life Sciences; Life Sciences, general; Biological Techniques; Analytical Chemistry; Microarrays; Computational Biology/Bioinformatics; Organic Chemistry
ISSN
1754-2189
eISSN
1750-2799
DOI
10.1038/nprot.2007.227
Publisher site
See Article on Publisher Site

Abstract

We present a detailed protocol for the structural analysis of protein-linked glycans. In this approach, appropriate for glycomics studies, N-linked glycans are released using peptide N-glycosidase F and O-linked glycans are released by reductive alkaline β-elimination. Using strategies based on mass spectrometry (matrix-assisted laser desorption/ionization–time of flight mass spectrometry and nano-electrospray ionization mass spectrometry/mass spectrometry (nano-ESI-MS-MS)), chemical derivatization, sequential exoglycosidase digestions and linkage analysis, the structures of the N- and/or O-glycans are defined. This approach can be used to study the glycosylation of isolated complex glycoproteins or of numerous glycoproteins encountered in a complex biological medium (cells, tissues and physiological fluids).

Journal

Nature ProtocolsSpringer Journals

Published: Jun 21, 2007

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