The Detection of Phospholipase-Resistant and -Sensitive Glycosyl-phosphatidylinositol Membrane Anchors by Western Blotting

M. L S Guther, M. L C Dealmeida, 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 (US)
Pages (from-to)249-255
Number of pages7
JournalAnalytical Biochemistry
Volume219
Issue number2
DOIs
StatePublished - Jun 1994
Externally publishedYes

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Glycosylphosphatidylinositols
Phospholipases
Anchors
Western Blotting
Membranes
Phosphoinositide Phospholipase C
Trypanosoma brucei brucei
Cell membranes
Blood Proteins
Epitopes
Membrane Proteins
Cell Membrane
Antibodies
Enzymes
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Biophysics
  • Biochemistry

Cite this

The Detection of Phospholipase-Resistant and -Sensitive Glycosyl-phosphatidylinositol Membrane Anchors by Western Blotting. / Guther, M. L S; Dealmeida, M. L C; Rosenberry, T. L.; Ferguson, M. A J.

In: Analytical Biochemistry, Vol. 219, No. 2, 06.1994, p. 249-255.

Research output: Contribution to journalArticle

Guther, M. L S ; Dealmeida, M. L C ; Rosenberry, T. L. ; Ferguson, M. A J. / The Detection of Phospholipase-Resistant and -Sensitive Glycosyl-phosphatidylinositol Membrane Anchors by Western Blotting. In: Analytical Biochemistry. 1994 ; Vol. 219, No. 2. pp. 249-255.
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