The Detection of phospholipase-resistant and -sensitive glycosyl-phosphatidylinositol membrane anchors by western blotting

Maria Lucia S. Guther, Maria Lucia Cardoso de Almeida, Terrone L. Rosenberry, Michael A.J. Ferguson

Research output: Contribution to journalArticle

26 Scopus citations

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 (US)
Pages (from-to)249-255
Number of pages7
JournalAnalytical Biochemistry
Volume219
Issue number2
DOIs
StatePublished - Jun 1994

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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