title: Glycopeptide enrichment using a combination of ZIC-HILIC and cotton wool for exploring the glycoproteome of wheat flour albumins.
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 (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.
authors.affiliation: Enzyme and Protein Chemistry, Søltofts Plads Building 224, Department of Systems Biology, Technical University of Denmark , DK-2800 Kongens Lyngby, Denmark.
authors.forename: Plaipol
authors.initials: P
authors.lastname: Dedvisitsakul
authors.forename: Susanne
authors.initials: S
authors.lastname: Jacobsen
authors.forename: Birte
authors.initials: B
authors.lastname: Svensson
authors.forename: Jakob
authors.initials: J
authors.lastname: Bunkenborg
authors.forename: Christine
authors.initials: C
authors.lastname: Finnie
authors.forename: Per
authors.initials: P
authors.lastname: Hägglund
mesh_terms: D000418:Albumins / Q000378:metabolism*; D000595:Amino Acid Sequence; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D032561:Cotton Fiber; D005433:Flour / Q000032:analysis; D006020:Glycopeptides / Q000737:chemistry / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D008969:Molecular Sequence Data; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism*; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry; D014908:Triticum / Q000378:metabolism*
chemical_list: D000418:Albumins; D006020:Glycopeptides; D006023:Glycoproteins; D010940:Plant Proteins; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase