THE DETECTION of PHOSPHOLIPASE-RESISTANT and PHOSPHOLIPASE-SENSITIVE GLYCOSYL-PHOSPHATIDYLINOSITOL MEMBRANE ANCHORS BY WESTERN BLOTTING
Rosenberry, T. L.
Is part ofAnalytical Biochemistry
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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. (C) 1994 Academic Press, Inc.
CitationAnalytical Biochemistry. San Diego: Academic Press Inc Jnl-comp Subscriptions, v. 219, n. 2, p. 249-255, 1994.
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