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

Z. Darula, Farkas Sarnyai, Katalin F. Medzihradszky

Research output: Contribution to journalArticle

14 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1-11
Number of pages11
JournalGlycoconjugate Journal
DOIs
Publication statusAccepted/In press - Jan 5 2016

Fingerprint

Glycosylation
Mucin-1
Glycopeptides
Mucins
Lectins
Bearings (structural)
Serum
Plant Lectins
Peptides
Wheat Germ Agglutinins
Workflow
Chromatography
Polysaccharides
Proteins
jacalin

Keywords

  • Automated glycopeptide identification
  • ETD
  • HCD
  • Human serum
  • O-glycosylation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

O-glycosylation sites identified from mucin core-1 type glycopeptides from human serum. / Darula, Z.; Sarnyai, Farkas; Medzihradszky, Katalin F.

In: Glycoconjugate Journal, 05.01.2016, p. 1-11.

Research output: Contribution to journalArticle

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