Cholera toxin binds to lipid rafts but has a limited specificity for ganglioside GM1

Lipid rafts and the formation of an immunological synapse are crucial for T‐cell activation. Binding of cholera toxin B subunit (CTB) to ganglioside GM1 is a marker to identify lipid rafts. Primary human T cells were isolated from healthy donors and were stimulated with superantigen staphylococcus e...

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Bibliographic Details
Published in:Immunology and cell biology 2007-07, Vol.85 (5), p.378-382
Main Authors: Blank, Norbert, Schiller, Martin, Krienke, Stefan, Wabnitz, Guido, Ho, Anthony D, Lorenz, Hanns‐Martin
Format: Article
Language:English
Subjects:
activation marker
Cell Proliferation - drug effects
Cholera Toxin - metabolism
Endocytosis - drug effects
Epitopes - metabolism
Fluorescein-5-isothiocyanate
G(M1) Ganglioside - metabolism
Glucosidases - metabolism
Humans
immunological synapse
Leukocytes, Mononuclear - drug effects
lipid raft
Lymphocyte Activation - drug effects
Membrane Microdomains - drug effects
Membrane Microdomains - metabolism
Mitogens - pharmacology
Protein Binding - drug effects
Staphylococcus
Substrate Specificity - drug effects
T-Lymphocytes - drug effects
T-Lymphocytes - metabolism
Trypsin - metabolism
T‐cell activation
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Summary:Lipid rafts and the formation of an immunological synapse are crucial for T‐cell activation. Binding of cholera toxin B subunit (CTB) to ganglioside GM1 is a marker to identify lipid rafts. Primary human T cells were isolated from healthy donors and were stimulated with superantigen staphylococcus enterotoxin B (SEB) and stained with cholera toxin B‐fluorescein isothiocyanate (CTB‐FITC). An optimized staining procedure is required to stain lipid rafts exclusively on the cell surface. Unstimulated T cells show a few CTB binding spots on the cell surface. The size and number of CTB‐binding lipid rafts are strongly upregulated during T‐cell activation in SEB‐stimulated CD4+ T cells. However, our data show that the specificity of CTB for GM1 ganglioside is limited, because the binding capacity is partly resistant to inhibition of ganglioside synthesis and sensitive to trypsin digestion. Our results indicate that the binding of FITC‐labeled CTB can be divided into at least three different categories: a specific binding of CTB to ganglioside GM1, a nonspecific binding of CTB probably to glycosylated surface proteins and a nonspecific binding of FITC to the cell surface.
ISSN:0818-9641
1440-1711
DOI:10.1038/sj.icb.7100045