Extended Sialylated O-Glycan Repertoire of Human Urinary Glycoproteins Discovered and Characterized Using Electron-Transfer/Higher-Energy Collision Dissociation
| Autor: | Zsuzsanna Darula, Katalin F. Medzihradszky, Adam Pap |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Proteomics Glycan Stereochemistry Peptide Biochemistry 03 medical and health sciences Residue (chemistry) Affinity chromatography Polysaccharides Tandem Mass Spectrometry Humans Glycoproteins chemistry.chemical_classification biology Chemistry Glycopeptides General Chemistry Glycopeptide Wheat germ agglutinin N-Acetylneuraminic Acid carbohydrates (lipids) 030104 developmental biology biology.protein Oxonium ion Glycoprotein Chromatography Liquid |
| Zdroj: | Journal of proteome research. 18(1) |
| ISSN: | 1535-3907 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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