CSC technology: selective labeling of glycoproteins by mild oxidation to phenotype cells

Methods Mol Biol. 2013:951:33-43. doi: 10.1007/978-1-62703-146-2_3.

Abstract

Cell surface glycoproteins represent important markers for the phenotyping of healthy and malignantly transformed cells. The mass spectrometry-based cell surface capturing (CSC) technology allows for extensive multiplexed identification and relative quantification of glycoproteins expressed on the cell surface at a given point in time. CSC technology is based on the mild oxidation of glycans from cell surface proteins on living cells. Oxidized glycans are tagged with a bifunctional linker molecule and glycopeptides are subsequently enriched by affinity chromatography. Here, we describe a step-by-step protocol of the CSC technology, which not only enables the identification of cell surface glycoproteins, but also the concurrent determination of protein N-glycosylation sites.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Biotinylation
  • Cell Membrane / metabolism
  • Chromatography, Liquid
  • Glycopeptides / chemistry
  • Glycopeptides / metabolism
  • Mass Spectrometry
  • Membrane Glycoproteins / analysis*
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / metabolism*
  • Phenotype
  • Proteolysis
  • Proteomics*
  • Salts / chemistry
  • Staining and Labeling / methods*

Substances

  • Glycopeptides
  • Membrane Glycoproteins
  • Salts