O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum

Darula, Zsuzsanna and Sarnyai, Farkas and Medzihradszky F., Katalin (2016) O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. GLYCOCONJUGATE JOURNAL, 33 (3). pp. 435-445. ISSN 0282-0080, ESSN: 1573-4986

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Abstract

In this work O-linked glycopeptides bearing mucin core-1 type structures were enriched from human serum. Since about 70 % of the O-glycans in human serum bind to the plant lectin Jacalin, we tested a previously successful protocol that combined Jacalin affinity enrichment on the protein- and peptide-level with ERLIC chromatography as a further enrichment step in between, to eliminate the high background of unmodified peptides. In parallel, we developed a simpler and significantly faster new workflow that used two lectins sequentially: wheat germ agglutinin and then Jacalin. The first lectin provides general glycopeptide enrichment, while the second specifically enriches O-linked glycopeptides with Galβ1-3GalNAcα structures. Mass spectrometric analysis of enriched samples showed that the new sample preparation method is more selective and sensitive than the former. Altogether, 52 unique glycosylation sites in 20 proteins were identified in this study. © 2015 Springer Science+Business Media New York

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