Mass-spectrometric identification and relative quantification of N-linked cell surface glycoproteins

TitleMass-spectrometric identification and relative quantification of N-linked cell surface glycoproteins
Publication TypeJournal Article
Year of Publication2009
AuthorsWollscheid B, Bausch-Fluck D, Henderson C, O'Brien R, Bibel M, Schiess R, Aebersold R, Watts J
JournalNat Biotechnol
Volume27
Pagination378-86
Date PublishedApr
PubMed Central IDPMC2829300
PMID19349973
Keywords*Algorithms, Amino Acid Sequence, Cell Membrane/*metabolism, Glycoproteins/*analysis/*chemistry, Mass Spectrometry/*methods, Molecular Sequence Data
AbstractAlthough 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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