Determination of glycosyl-phosphatidylinositol membrane protein anchorage.
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.
Hooper
N M
NM
School of Biochemistry and Molecular Biology, University of Leeds, Leeds LS2 9JT. n.m.hooper@leeds.ac.uk
D018076:DNA, Complementary; D003902:Detergents / Q000737:chemistry; D017261:Glycosylphosphatidylinositols; D008055:Lipids / Q000737:chemistry*; D012995:Solubility
D018076:DNA, Complementary; D003902:Detergents; D017261:Glycosylphosphatidylinositols; D008055:Lipids