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

Darula, Zsuzsanna and Pap, Ádám and Medzihradszky F., Katalin (2019) Extended Sialylated O-Glycan Repertoire of Human Urinary Glycoproteins Discovered and Characterized Using Electron-Transfer/Higher-Energy Collision Dissociation. JOURNAL OF PROTEOME RESEARCH, 18 (1). pp. 280-291. ISSN 1535-3893

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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 Lewis(X/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.

Item Type: Article
Additional Information: Funding Agency and Grant Number: Ministry for National Economy [GOP-1.1.1-11-2012-0452, GINOP-2.3.2-15-2016-00001, GINOP-2.3.2-15-2016-00020] Funding text: The authors wish to thank Robert Chalkley for his constructive comments on the discussion section, and Kirk Hansen for correcting our Hunglish mistakes. We thank the MTA Cloud (https://cloud.mta.hu/) for housing our Protein Prospector server. This work was supported by the following grants: New Szechenyi Plan GOP-1.1.1-11-2012-0452, the Economic Development and Innovation Operative Programmes GINOP-2.3.2-15-2016-00001, and GINOP-2.3.2-15-2016-00020 from the Ministry for National Economy.
Uncontrolled Keywords: EXPRESSION; BINDING; DIFFERENTIATION; PEPTIDE; IDENTIFICATION; Mass Spectrometry; Acetylation; glycosylation; affinity chromatography; EThcD; blood-type antigen; disialic acid; O-acetyl-sialic acid; O-glycopeptides; urinary glycoproteins; SIALIC-ACID; LINKED OLIGOSACCHARIDES;
Subjects: Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia > QH3011 Biochemistry / biokémia
SWORD Depositor: MTMT SWORD
Depositing User: MTMT SWORD
Date Deposited: 21 Nov 2019 10:25
Last Modified: 21 Nov 2019 10:25
URI: http://real.mtak.hu/id/eprint/103474

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