The detection of phospholipase-resistant and -sensitive glycosyl-phosphatidylinositol membrane anchors by Western blotting

TY - JOUR

T1 - The detection of phospholipase-resistant and -sensitive glycosyl-phosphatidylinositol membrane anchors by Western blotting

AU - Guther, M. L. S.

AU - de Almeida, M. L. C.

AU - Rosenberry, T. L.

AU - Ferguson, M. A. J.

PY - 1994

Y1 - 1994

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.

U2 - 10.1006/abio.1994.1264

DO - 10.1006/abio.1994.1264

M3 - Article

C2 - 7521581

SN - 0003-2697

VL - 219

SP - 249

EP - 255

JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 2

ER -