Glycopeptide Enrichment Using a Combination of ZIC-HILIC and Cotton Wool for Exploring the Glycoproteome of Wheat Flour Albumins
Copyright policy )Glycopeptide Enrichment Using a Combination of ZIC-HILIC and Cotton Wool for Exploring the Glycoproteome of Wheat Flour Albumins
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.
- Technical University of Denmark Denmark
- University of Copenhagen Denmark
- University of Southern Denmark Denmark
- Copenhagen University Hospital Denmark
- University of Copenhagen Denmark
Glycosylation, Glycopeptide enrichment, Liquid/instrumentation, Flour, Molecular Sequence Data, Triticum/metabolism, deglycosylation, N-glycosylation, Glycoproteins/metabolism, cotton, Glycopeptides/chemistry, Flour/analysis, Tandem Mass Spectrometry, Albumins, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase, isotope labeling, Amino Acid Sequence, Cotton Fiber, HILIC, cereal proteomics, Triticum, mass spectrometry, Plant Proteins/metabolism, Glycoproteins, Plant Proteins, Chromatography, Solid Phase Extraction, Glycopeptides, deamidation, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase/metabolism, Albumins/metabolism, Chromatography, Liquid
Glycosylation, Glycopeptide enrichment, Liquid/instrumentation, Flour, Molecular Sequence Data, Triticum/metabolism, deglycosylation, N-glycosylation, Glycoproteins/metabolism, cotton, Glycopeptides/chemistry, Flour/analysis, Tandem Mass Spectrometry, Albumins, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase, isotope labeling, Amino Acid Sequence, Cotton Fiber, HILIC, cereal proteomics, Triticum, mass spectrometry, Plant Proteins/metabolism, Glycoproteins, Plant Proteins, Chromatography, Solid Phase Extraction, Glycopeptides, deamidation, Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase/metabolism, Albumins/metabolism, Chromatography, Liquid
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