Glycoproteomic analysis by two-dimensional electrophoresis

Methods Mol Biol. 2009:520:59-74. doi: 10.1007/978-1-60327-811-9_5.

Abstract

Changes in N-linked glycosylation are known to occur during the development of cancer. For example, increased branching of oligosaccharides has been associated with metastasis and has been correlated to tumor progression in human cancers of the breast, colon, and melanomas. Increases in core fucosylation have also been associated with the development of hepatocellular carcinoma (HCC). To a large extent, the proteins to which these N-linked glycans are attached have been unknown. However, with the advent of sensitive glycan analysis and proteomic technologies, the ability to comprehensively identify all the fucosylated proteins in a given population is now a possibility. This method, generally referred to as targeted glycoproteomics, is shown as applied to the detection of proteins present in the fucosylated proteome of a liver cancer cell line but is generally enough to be applied in many other situations.

MeSH terms

  • Antibodies
  • Cell Line, Tumor
  • Cells, Cultured
  • Electrophoresis, Gel, Two-Dimensional
  • Enzyme-Linked Immunosorbent Assay
  • Fluorescent Dyes
  • Fucose / metabolism
  • Glycoproteins / analysis*
  • Glycosylation
  • Humans
  • Lectins / chemistry
  • Lectins / isolation & purification
  • Oxidation-Reduction
  • Protein Conformation
  • Proteomics / methods*

Substances

  • Antibodies
  • Fluorescent Dyes
  • Glycoproteins
  • Lectins
  • Fucose