Determination of glycosyl-phosphatidylinositol membrane protein anchorage

Proteomics. 2001 Jun;1(6):748-55. doi: 10.1002/1615-9861(200106)1:6<748::AID-PROT748>3.0.CO;2-T.

Abstract

A diverse range of proteins are modified by the post-translational covalent attachment of a glycosyl-phosphatidylinositol (GPI) membrane anchor. Hydropathy plots and other computer algorithms can be used to predict the presence of a GPI anchor attachment signal in the nascent polypeptide chain. However, the presence of a GPI anchor on the mature protein requires experimental evidence, including sensitivity of the protein to release from cells or membranes with bacterial phosphatidylinositol-specific phospholipase C, recognition by anti-cross-reacting determinant antibodies, or metabolic labelling with components of the anchor. GPI-anchored proteins are resistant to solubilisation with detergents such as Triton X-100 due to their association with cholesterol and glycosphingolipids in membrane domains known as lipid rafts. This detergent insolubility can be used to provide evidence for the presence of a GPI anchor on a protein and to isolate lipid rafts.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • DNA, Complementary
  • Detergents / chemistry
  • Glycosylphosphatidylinositols*
  • Lipids / chemistry*
  • Solubility

Substances

  • DNA, Complementary
  • Detergents
  • Glycosylphosphatidylinositols
  • Lipids