Glycopeptide Enrichment Using a Combination of ZIC-HILIC and Cotton Wool for Exploring the Glycoproteome of Wheat Flour Albumins

Plaipol Dedvisitsakul, Susanne Jacobsen, Birte Svensson, Jakob Bunkenborg, Christine Finnie, Per Hägglund

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    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 18O labeling and deglycosylation using peptide-N-glycosidase A (PNGase A) in H218O. 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
    Pages (from-to)2696-2703
    ISSN1535-3893
    DOIs
    Publication statusPublished - 2014

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