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Region Hovedstaden - en del af Københavns Universitetshospital
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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.

OriginalsprogEngelsk
TidsskriftJournal of Proteome Research
Vol/bind13
Udgave nummer5
Sider (fra-til)2696-703
Antal sider8
ISSN1535-3893
DOI
StatusUdgivet - 2 maj 2014
Eksternt udgivetJa

ID: 44805445