Glycan side reaction may compromise ETD-based Glycopeptide identification

Z. Darula, Katalin F. Medzihradszky

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Tris(hydroxymethyl)aminomethane (Tris) is one of the most frequently used buffer ingredients. Among other things, it is recommended and is usually used for lectin-based affinity enrichment of glycopeptides. Here we report that sialic acid, a common 'capping' unit in both N- and O-linked glycans may react with this chemical, and this side reaction may compromise glycopeptide identification when ETD spectra are the only MS/MS data used in the database search. We show that the modification may alter N- as well as O-linked glycans, the Tris-derivative is still prone to fragmentation both in 'beam-type' CID (HCD) and ETD experiments, at the same time - since the acidic carboxyl group was 'neutralized' - it will display a different retention time than its unmodified counterpart. We also suggest solutions that - when incorporated into existing search engines - may significantly improve the reliability of glycopeptide assignments. [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)977-987
Number of pages11
JournalJournal of the American Society for Mass Spectrometry
Volume25
Issue number6
DOIs
Publication statusPublished - 2014

Fingerprint

Glycopeptides
Polysaccharides
Search Engine
Tromethamine
N-Acetylneuraminic Acid
Search engines
Lectins
Buffers
Databases
Derivatives
Experiments

Keywords

  • CID
  • ETD
  • Glycopeptide
  • HCD
  • Sialic acid
  • Side reaction

ASJC Scopus subject areas

  • Structural Biology
  • Spectroscopy
  • Medicine(all)

Cite this

Glycan side reaction may compromise ETD-based Glycopeptide identification. / Darula, Z.; Medzihradszky, Katalin F.

In: Journal of the American Society for Mass Spectrometry, Vol. 25, No. 6, 2014, p. 977-987.

Research output: Contribution to journalArticle

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