Glycopeptide enrichment using a combination of ZIC-HILIC and cotton wool for exploring the glycoproteome of wheat flour albumins
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Glycopeptide enrichment using a combination of ZIC-HILIC and cotton wool for exploring the glycoproteome of wheat flour albumins. / Dedvisitsakul, Plaipol; Jacobsen, Susanne; Svensson, Birte; Bunkenborg, Jakob; Finnie, Christine; Hägglund, Per.
In: Journal of Proteome Research, Vol. 13, No. 5, 02.05.2014, p. 2696-2703.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Glycopeptide enrichment using a combination of ZIC-HILIC and cotton wool for exploring the glycoproteome of wheat flour albumins
AU - Dedvisitsakul, Plaipol
AU - Jacobsen, Susanne
AU - Svensson, Birte
AU - Bunkenborg, Jakob
AU - Finnie, Christine
AU - Hägglund, Per
PY - 2014/5/2
Y1 - 2014/5/2
N2 - 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.
AB - 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.
KW - cereal proteomics
KW - cotton
KW - deamidation
KW - deglycosylation
KW - Glycopeptide enrichment
KW - HILIC
KW - isotope labeling
KW - mass spectrometry
KW - N-glycosylation
U2 - 10.1021/pr401282r
DO - 10.1021/pr401282r
M3 - Journal article
C2 - 24654981
AN - SCOPUS:84899882471
VL - 13
SP - 2696
EP - 2703
JO - Journal of Proteome Research
JF - Journal of Proteome Research
SN - 1535-3893
IS - 5
ER -
ID: 240159661