Title : Affinity Purification of Glycosylphosphatidylinositol-anchored Proteins by Alpha-Toxin - Huang_2022_Methods.Mol.Biol_2303_251 |
Author(s) : Huang K , Park S |
Ref : Methods Mol Biol , 2303 :251 , 2022 |
Abstract :
The glycosylphosphatidylinositol (GPI)-anchor modification attaches a lipid anchor to the C-terminus of a protein, tethering the protein to the cell surface membrane. From this membrane-bound state, GPI-anchored proteins (GPI-APs) can be released into the extracellular space by multiple mechanisms, including proteolytic shedding and GPI lipase activity. Since the core GPI structure is co-released with the protein by GPI lipase activity, while removed from the protein by proteolytic cleavage, affinity purification by alpha-toxin (alphaToxin), which binds to the core domain of the GPI-anchor, isolates GPI-containing proteins from the culture medium. The following method details a technique for affinity purification of GP-APs using His-tagged alphaToxin for identification of GPI-anchored proteins, analysis of the GPI-anchor status of a protein of interest, or purification for subsequent biochemical analysis. |
PubMedSearch : Huang_2022_Methods.Mol.Biol_2303_251 |
PubMedID: 34626384 |
Huang K, Park S (2022)
Affinity Purification of Glycosylphosphatidylinositol-anchored Proteins by Alpha-Toxin
Methods Mol Biol
2303 :251
Huang K, Park S (2022)
Methods Mol Biol
2303 :251