Premium
Revealing the glycation sites in synthetic neoglycoconjugates formed by conjugation of the antigenic monosaccharide hapten of Vibrio cholerae O1, serotype Ogawa with the BSA protein carrier using LC‐ESI‐QqTOF‐MS/MS
Author(s) -
Jahouh Farid,
Saksena Rina,
Kováč Pavol,
Banoub Joseph
Publication year - 2012
Publication title -
journal of mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 121
eISSN - 1096-9888
pISSN - 1076-5174
DOI - 10.1002/jms.2974
Subject(s) - chemistry , hapten , matrix assisted laser desorption/ionization , chromatography , glycation , vibrio cholerae , electrospray ionization , lysine , tandem mass spectrometry , mass spectrometry , stereochemistry , biochemistry , organic chemistry , antigen , bacteria , amino acid , desorption , genetics , receptor , adsorption , biology
In this manuscript, we present the determination of glycation sites in synthetic neoglycoconjugates formed by conjugation of the antigenic monosaccharide hapten of Vibrio cholerae O1 serotype Ogawa to BSA using nano‐ liquid chromatography electrospray ionization quadrupole time‐of‐flight tandem mass spectroscopy (LC‐ESI‐QqTOF‐MS/MS). The matrix‐assisted laser desorption/ionization‐TOF/TOF‐MS/MS analyses of the tryptic digests of the glycoconjugates having a hapten:BSA ratio of 4.3:1, 6.6:1 and 13.2:1 revealed only three glycation sites, on the following lysine residues: Lys 235, Lys 437 and Lys 455. Digestion of the neoglycoconjugates with the proteases trypsin and GluC V8 gave complementary structural information and was shown to maximize the number of recognized glycation sites. Here, we report identification of 20, 27 and 33 glycation sites using LC‐ESI‐QqTOF‐MS/MS analysis of a series of synthetic neoglycoconjugates with a hapten:BSA ratio of, respectively, 4.3:1, 6.6:1 and 13.2:1. We also tentatively propose that all the glycated lysine residues are located mainly near the outer surface of the protein. Copyright © 2012 John Wiley & Sons, Ltd.