TY - JOUR
T1 - The Detection of phospholipase-resistant and -sensitive glycosyl-phosphatidylinositol membrane anchors by western blotting
AU - Guther, Maria Lucia S.
AU - de Almeida, Maria Lucia Cardoso
AU - Rosenberry, Terrone L.
AU - Ferguson, Michael A.J.
PY - 1994/6
Y1 - 1994/6
N2 - Glycosyl-phosphatidylinositol (GPI) membrane anchors are present on a large number of eukaryotic plasma membrane proteins. Some of these anchors can be cleaved with bacterial phosphatidylinositol-specific phospholipases C, and a glycosyl-phosphatidylinositol-specific phospholipase C from Trypanosoma brucei, to reveal an epitope called the cross-reacting determinant. Other glycosyl-phosphatidylinositol anchors are resistant to the action of these enzymes prior to treatment with mild base. A simple method is described for identifying both phospholipase-sensitive and -resistant anchors using anti-cross-reacting determinant antibodies on Western blots. This procedure represents a high-sensitivity general method for the identification of GPI-anchored proteins.
AB - Glycosyl-phosphatidylinositol (GPI) membrane anchors are present on a large number of eukaryotic plasma membrane proteins. Some of these anchors can be cleaved with bacterial phosphatidylinositol-specific phospholipases C, and a glycosyl-phosphatidylinositol-specific phospholipase C from Trypanosoma brucei, to reveal an epitope called the cross-reacting determinant. Other glycosyl-phosphatidylinositol anchors are resistant to the action of these enzymes prior to treatment with mild base. A simple method is described for identifying both phospholipase-sensitive and -resistant anchors using anti-cross-reacting determinant antibodies on Western blots. This procedure represents a high-sensitivity general method for the identification of GPI-anchored proteins.
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U2 - 10.1006/abio.1994.1264
DO - 10.1006/abio.1994.1264
M3 - Article
C2 - 7521581
AN - SCOPUS:0028276578
SN - 0003-2697
VL - 219
SP - 249
EP - 255
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -