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

M. L. S. Guther, M. L. C. de Almeida, T. L. Rosenberry, M. A. J. Ferguson

    Research output: Contribution to journalArticle

    26 Citations (Scopus)

    Abstract

    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.
    Original languageEnglish
    Pages (from-to)249-255
    Number of pages7
    JournalAnalytical Biochemistry
    Volume219
    Issue number2
    DOIs
    Publication statusPublished - 1994

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