Extended Sialylated O-Glycan Repertoire of Human Urinary Glycoproteins Discovered and Characterized Using Electron-Transfer/Higher-Energy Collision Dissociation

Z. Darula, Ádám Pap, Katalin F. Medzihradszky

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

A relatively novel activation technique, electron-transfer/higher-energy collision dissociation (EThcD) was used in the LC-MS/MS analysis of tryptic glycopeptides enriched with wheat germ agglutinin from human urine samples. We focused on the characterization of mucin-type O-glycopeptides. EThcD in a single spectrum provided information on both the peptide modified and the glycan carried. Unexpectedly, glycan oxonium ions indicated the presence of O-acetyl, and even O-diacetyl-sialic acids. B and Y fragment ions revealed that (i) in core 1 structures the Gal residue featured the O-acetyl-sialic acid, when there was only one in the glycan; (ii) several glycopeptides featured core 1 glycans with disialic acids, in certain instances O-acetylated; (iii) the disialic acid was linked to the GalNAc residue whatever the degree of O-acetylation; (iv) core 2 isomers with a single O-acetyl-sialic acid were chromatographically resolved. Glycan fragmentation also helped to decipher additional core 2 oligosaccharides: a LacdiNAc-like structure, glycans carrying sialyl LewisX/A at different stages of O-acetylation, and blood antigens. A sialo core 3 structure was also identified. We believe this is the first study when such structures were characterized from a very complex mixture and were linked not only to a specific protein, but also the sites of modifications have been determined.

Original languageEnglish
Pages (from-to)280-291
Number of pages12
JournalJournal of Proteome Research
Volume18
Issue number1
DOIs
Publication statusPublished - Jan 4 2019

Fingerprint

Energy Transfer
Polysaccharides
Glycoproteins
Electrons
Glycopeptides
Acetylation
Ions
Sialic Acids
Diacetyl
Wheat Germ Agglutinins
Acids
Mucins
Complex Mixtures
Oligosaccharides
Isomers
Blood
Chemical activation
Urine
Antigens
Peptides

Keywords

  • affinity chromatography
  • blood-type antigen
  • disialic acid
  • EThcD
  • mass spectrometry
  • O-acetyl-sialic acid
  • O-glycopeptides
  • urinary glycoproteins
  • wheat germ agglutinin

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Extended Sialylated O-Glycan Repertoire of Human Urinary Glycoproteins Discovered and Characterized Using Electron-Transfer/Higher-Energy Collision Dissociation. / Darula, Z.; Pap, Ádám; Medzihradszky, Katalin F.

In: Journal of Proteome Research, Vol. 18, No. 1, 04.01.2019, p. 280-291.

Research output: Contribution to journalArticle

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