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Glycopeptide enrichment using a combination of ZIC-HILIC and cotton wool for exploring the glycoproteome of wheat flour albumins

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  • Plaipol Dedvisitsakul
  • Susanne Jacobsen
  • Birte Svensson
  • Jakob Bunkenborg
  • Christine Finnie
  • Per Hägglund
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Hydrophilic liquid chromatography (HILIC) is used extensively as a sample preparation step for glycopeptide enrichment in proteome research. Here, we have applied cotton wool and a zwitterionic HILIC (ZIC-HILIC) resin in solid-phase extraction microcolumns to provide a higher loading capacity and broader specificity for glycopeptide enrichment. This strategy was applied to tryptic digests of wheat flour albumin extracts followed by simulataneous site-specific (18)O labeling and deglycosylation using peptide-N-glycosidase A (PNGase A) in H(2)(18)O. Subsequent LC-MS/MS analysis allowed for assignment of 78 N-glycosylation sites in 67 albumin proteins. Bioinformatic analysis revealed that several of the identified glycoproteins show sequence similarity to known food allergens. In addition, the potential impact of some of the identified glycoproteins on wheat beer quality is discussed.

Original languageEnglish
JournalJournal of Proteome Research
Volume13
Issue number5
Pages (from-to)2696-703
Number of pages8
ISSN1535-3893
DOIs
Publication statusPublished - 2 May 2014
Externally publishedYes

ID: 44805445