Method for investigation of oligosaccharides from glycopeptides: direct determination of glycosylation sites in proteins

Analytical Chemistry
Erika LattovaHélène Perreault

Abstract

Characterization of glycopeptides has become an important tool toward a better understanding of the molecular details in carbohydrate-protein interactions. In this approach, oligosaccharides are commonly not detectable under mass spectrometric conditions because of ionization suppression by deglycosylated peptides. Their composition is only deduced from the mass differences between glycopeptides and corresponding deglycosylated peptides. Here, we describe how carbohydrates can be easily detected in the PNGase-treated samples and structurally investigated next to the peptides. The efficacy of this method is demonstrated through the analysis of tryptic glycopeptides obtained from human IgG. Following deglycosylation with PNGaseF and derivatization with phenylhydrazine, MALDI spectra produced ion peaks of labeled oligosaccharides and deglycosylated peptides. The relative abundances of individual oligosaccharides were consistent with those of the glycopeptides. MALDI-MS/MS provided useful data for the structural elucidation of oligosaccharides, including the assignment of dominant isomers and glycosylation sites in peptides. MALDI-MS/MS fragmentation patterns of deglycosylated peptide ions indicated glycosylation sites at asparagin...Continue Reading

References

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Jul 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Erika Lattova, Hélène Perreault
May 3, 2005·Journal of the American Society for Mass Spectrometry·Erika LattováOleg Krokhin

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Citations

Mar 21, 2008·Journal of the American Society for Mass Spectrometry·Erika LattováHélène Perreault
May 26, 2011·Molecular & Cellular Proteomics : MCP·Erika LattováBoguslaw Tomanek
Mar 9, 2011·Journal of Pharmaceutical and Biomedical Analysis·Keita Yamada, Kazuaki Kakehi
Mar 30, 2010·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Dorota BartusikGino Fallone
Aug 28, 2009·Journal of Pharmaceutical and Biomedical Analysis·Dorota BartusikGino Fallone
Jan 25, 2013·Mass Spectrometry Reviews·Erika Lattová, Hélène Perreault
Jul 28, 2016·Journal of Pharmaceutical and Biomedical Analysis·Sachio YamamotoShigeo Suzuki
May 16, 2007·Electrophoresis·Haleem J Issaq, Timothy D Veenstra
Apr 25, 2020·Mass Spectrometry Reviews·David J Harvey
Oct 12, 2013·Analytical Chemistry·Edward D Bodnar, Hélène Perreault
Dec 30, 2006·Analytical Chemistry·Caterina TemporiniGabriella Massolini
Jul 20, 2007·Chemical Reviews·Haleem J IssaqTimothy D Veenstra

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