Mass-spectrometric identification and relative quantification of N-linked cell surface glycoproteins
Open accessAbstract
Although the classification of cell types often relies on the identification of cell surface proteins as differentiation markers, flow cytometry requires suitable antibodies and currently permits detection of only up to a dozen differentiation markers in a single measurement. We use multiplexed mass-spectrometric identification of several hundred N-linked glycosylation sites specifically from cell surface-exposed glycoproteins to phenotype cells without antibodies in an unbiased fashion and without a priori knowledge. We apply our cell surface-capturing (CSC) technology, which covalently labels extracellular glycan moieties on live cells, to the detection and relative quantitative comparison of the cell surface N-glycoproteomes of T and B cells, as well as to monitor changes in the abundance of cell surface N-glycoprotein markers during T-cell activation and the controlled differentiation of embryonic stem cells into the neural lineage. A snapshot view of the cell surface N-glycoproteins will enable detection of panels of N-glycoproteins as potential differentiation markers that are currently not accessible by other means.
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PM enrichment by chemical derivatization and affinity purification lowers organelle contamination and improves detection of low-abundance membrane receptors.
“To profile cell surface proteins by MS, PM enrichment by chemical derivatization–affinity purification confers reduced organelle contamination and improved detection of low-abundance membrane receptors (107, 108).”
From The Impact of Mass Spectrometry–Based Proteomics on Fundamental Discoveries in Virology (2014) · cites this paper as [108]
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- Authors
- Wollscheid, B., Bausch-Fluck, D., Henderson, C., O'Brien, R., Bibel, M., Schiess, R., Aebersold, R., & Watts, J. D.
- Affiliations
- Institute for Systems Biology (US)
- Novartis Institutes for BioMedical Research (CH)
- University of Zurich (CH)
- Corresponding author
- Wollscheid, Bernd
- Venue
- Nature Biotechnology
- Published
- 2009-04
- Volume
- 27
- Issue
- 4
- Pages
- 378-386
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC2829300
- OpenAlex
- W2081927805
- Cited by
- 547
- References
- 48
- Citations by year
- 2026: 16 · 2025: 15 · 2024: 28 · 2023: 30 · 2022: 32 · 2021: 40 · 2020: 25 · 2019: 27 · 2018: 23 · 2017: 28 · 2016: 33 · 2015: 33 · 2014: 36 · 2013: 46 · 2012: 51
- FWCI
- 17.97
- Type
- journal article
- Language
- en
- Update notices
- erratum (2009-09-01)
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Canary Foundation
- China Scholarship Council
- National Institutes of Health (n01hv28179-12-0-0)
- University of California, San Francisco
- National Heart, Lung, and Blood Institute (N01 HV028179, N01-HV-28179, N01-HV-28179-22)
- National Institute of Allergy and Infectious Diseases (R01-AI-51344-01, R01AI051344)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Glycosylation and Glycoproteins Research
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Keywords
- N-linked glycoproteins, N-glycosylation sites, T cell activation, embryonic stem cell differentiation, neural progenitor cells, differentiation markers, B cells
- MeSH terms
- Algorithms, Amino Acid Sequence, Cell Membrane, Glycoproteins, Molecular Sequence Data, Mass Spectrometry
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