A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation. / Hägglund, Per; Bunkenborg, Jakob; Elortza, Felix; Jensen, Ole Nørregaard; Roepstorff, Peter.

In: Journal of Proteome Research, Vol. 3, No. 3, 01.05.2004, p. 556-566.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Hägglund, P, Bunkenborg, J, Elortza, F, Jensen, ON & Roepstorff, P 2004, 'A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation', Journal of Proteome Research, vol. 3, no. 3, pp. 556-566. https://doi.org/10.1021/pr034112b

APA

Hägglund, P., Bunkenborg, J., Elortza, F., Jensen, O. N., & Roepstorff, P. (2004). A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation. Journal of Proteome Research, 3(3), 556-566. https://doi.org/10.1021/pr034112b

Vancouver

Hägglund P, Bunkenborg J, Elortza F, Jensen ON, Roepstorff P. A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation. Journal of Proteome Research. 2004 May 1;3(3):556-566. https://doi.org/10.1021/pr034112b

Author

Hägglund, Per ; Bunkenborg, Jakob ; Elortza, Felix ; Jensen, Ole Nørregaard ; Roepstorff, Peter. / A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation. In: Journal of Proteome Research. 2004 ; Vol. 3, No. 3. pp. 556-566.

Bibtex

@article{4bf7090fb3794ad5a7386972ad98821c,
title = "A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation",
abstract = "Characterization of glycoproteins using mass spectrometry ranges from determination of carbohydrate-protein linkages to the full characterization of all glycan structures attached to each glycosylation site. In a novel approach to identify N-glycosylation sites in complex biological samples, we performed an enrichment of glycosylated peptides through hydrophilic interaction liquid chromatography (HILIC) followed by partial deglycosylation using a combination of endo-β-N-acetylglucosaminidases (EC 3.2.1.96). After hydrolysis with these enzymes, a single N-acetylglucosamine (GlcNAc) residue remains linked to the asparagine residue. The removal of the major part of the glycan simplifies the MS/MS fragment ion spectra of glycopeptides, while the remaining GlcNAc residue enables unambiguous assignment of the glycosylation site together with the amino acid sequence. We first tested our approach on a mixture of known glycoproteins, and subsequently the method was applied to samples of human plasma obtained by lectin chromatography followed by 1D gel-electrophoresis for determination of 62 glycosylation sites in 37 glycoproteins.",
keywords = "Endoglycosidase, Glycosylation, HILIC, Lectin affinity chromatography, Mass spectrometry, Plasma proteins, Post-translational modifications, Proteomics",
author = "Per H{\"a}gglund and Jakob Bunkenborg and Felix Elortza and Jensen, {Ole N{\o}rregaard} and Peter Roepstorff",
year = "2004",
month = "5",
day = "1",
doi = "10.1021/pr034112b",
language = "English",
volume = "3",
pages = "556--566",
journal = "Journal of Proteome Research",
issn = "1535-3893",
publisher = "American Chemical Society",
number = "3",

}

RIS

TY - JOUR

T1 - A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation

AU - Hägglund, Per

AU - Bunkenborg, Jakob

AU - Elortza, Felix

AU - Jensen, Ole Nørregaard

AU - Roepstorff, Peter

PY - 2004/5/1

Y1 - 2004/5/1

N2 - Characterization of glycoproteins using mass spectrometry ranges from determination of carbohydrate-protein linkages to the full characterization of all glycan structures attached to each glycosylation site. In a novel approach to identify N-glycosylation sites in complex biological samples, we performed an enrichment of glycosylated peptides through hydrophilic interaction liquid chromatography (HILIC) followed by partial deglycosylation using a combination of endo-β-N-acetylglucosaminidases (EC 3.2.1.96). After hydrolysis with these enzymes, a single N-acetylglucosamine (GlcNAc) residue remains linked to the asparagine residue. The removal of the major part of the glycan simplifies the MS/MS fragment ion spectra of glycopeptides, while the remaining GlcNAc residue enables unambiguous assignment of the glycosylation site together with the amino acid sequence. We first tested our approach on a mixture of known glycoproteins, and subsequently the method was applied to samples of human plasma obtained by lectin chromatography followed by 1D gel-electrophoresis for determination of 62 glycosylation sites in 37 glycoproteins.

AB - Characterization of glycoproteins using mass spectrometry ranges from determination of carbohydrate-protein linkages to the full characterization of all glycan structures attached to each glycosylation site. In a novel approach to identify N-glycosylation sites in complex biological samples, we performed an enrichment of glycosylated peptides through hydrophilic interaction liquid chromatography (HILIC) followed by partial deglycosylation using a combination of endo-β-N-acetylglucosaminidases (EC 3.2.1.96). After hydrolysis with these enzymes, a single N-acetylglucosamine (GlcNAc) residue remains linked to the asparagine residue. The removal of the major part of the glycan simplifies the MS/MS fragment ion spectra of glycopeptides, while the remaining GlcNAc residue enables unambiguous assignment of the glycosylation site together with the amino acid sequence. We first tested our approach on a mixture of known glycoproteins, and subsequently the method was applied to samples of human plasma obtained by lectin chromatography followed by 1D gel-electrophoresis for determination of 62 glycosylation sites in 37 glycoproteins.

KW - Endoglycosidase

KW - Glycosylation

KW - HILIC

KW - Lectin affinity chromatography

KW - Mass spectrometry

KW - Plasma proteins

KW - Post-translational modifications

KW - Proteomics

UR - http://www.scopus.com/inward/record.url?scp=4444269089&partnerID=8YFLogxK

U2 - 10.1021/pr034112b

DO - 10.1021/pr034112b

M3 - Journal article

C2 - 15253437

AN - SCOPUS:4444269089

VL - 3

SP - 556

EP - 566

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

IS - 3

ER -

ID: 240161809