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Matrix‐assisted laser desorption/ionization on‐target method for the investigation of oligosaccharides and glycosylation sites in glycopeptides and glycoproteins
Author(s) -
Lattová Erika,
Chen Vincent C.,
Varma Sonal,
Bezabeh Tedros,
Perreault Hélène
Publication year - 2007
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3007
Subject(s) - chemistry , glycopeptide , glycoprotein , chromatography , matrix assisted laser desorption/ionization , glycosylation , glycan , peptide , mass spectrometry , oligosaccharide , matrix (chemical analysis) , biochemistry , desorption , organic chemistry , adsorption , antibiotics
The significance of glycoproteins in living systems instigates the ceaseless expansion of new techniques and procedures for the analysis of biological samples. Many of these applications are focused on improving the detection limit of analyzed material. In a previous study, we described a procedure for the detection of oligosaccharides cleaved from tryptic glycopeptides. Treatment of deglycosylated fractions with phenylhydrazine gave rise to peaks consistent with labeled glycans, and both types of compounds—deglycosylated peptides and oligosaccharides—were recorded from one spot and observed in one matrix‐assisted laser desorption/ionization (MALDI) mass spectrum for the first time. Here, we added an additional step to this simple procedure of deglycosylating glycopeptides directly from the target spot of the first analyzed glycosylated peptides. For the purpose of this new study, a mixture of 2‐aza‐2‐thiothymine and phenylhydrazine hydrochloride showed to be an excellent matrix for glycopeptides, oligosaccharides, deglycosylated peptides and moreover it allowed PNGaseF to be active enough to cleave oligosaccharides from peptides. The efficiency of this procedure is demonstrated on a series of intact glycoproteins and on the analysis of tryptic peptides obtained from IgG and total mouse serum. This one‐step on‐target deglycosylation method with subsequent derivatization on the same spot makes MALDI‐MS analyses of glycopeptides fast, simple and accessible for biological samples, where classical procedures cannot produce useful results. Copyright © 2007 John Wiley & Sons, Ltd.