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Stable immobilization of lipid vesicles for kinetic studies using surface plasmon resonance.
- Source :
-
Analytical biochemistry [Anal Biochem] 1994 May 01; Vol. 218 (2), pp. 405-12. - Publication Year :
- 1994
-
Abstract
- In order to study the kinetics of binding between membrane vesicle surface receptors to the lepidopteran insecticidal toxins from Bacillus thuringiensis using surface plasmon resonance, we have developed a technique to immobilize membrane vesicles purified from the brush border of dissected guts from the lepidopteran insect pest Choristoneura fumiferana. Two methods using immobilized immunoglobulins against either avidin or biotin were successful in achieving stable immobilization of the vesicles (> 1.5 h). Specificity of the immobilized receptors exposed on the vesicle surface was demonstrated, in part, by the inability of bovine serum albumin to bind to the immobilized brush border membrane vesicles. Homologous and heterologous competition experiments further demonstrated specific binding of trypsin-activated CryIA(c) toxin to the cell-surface receptors on the vesicles. Kinetic rate constants for activated cryIA(b) toxin binding to brush border vesicles were determined, revealing the presence of a high-affinity receptor on the surface of the immobilized brush border membrane vesicles.
- Subjects :
- Animals
Arthropod Venoms metabolism
Bacillus thuringiensis
Bacillus thuringiensis Toxins
Bacterial Proteins toxicity
Bacterial Toxins toxicity
Biosensing Techniques
Drug Stability
Endotoxins toxicity
Hemolysin Proteins
Kinetics
Lipids chemistry
Membranes metabolism
Microvilli metabolism
Models, Biological
Pest Control, Biological
Substrate Specificity
Bacterial Proteins metabolism
Bacterial Toxins metabolism
Endotoxins metabolism
Lipid Metabolism
Liposomes
Subjects
Details
- Language :
- English
- ISSN :
- 0003-2697
- Volume :
- 218
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Analytical biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- 8074300
- Full Text :
- https://doi.org/10.1006/abio.1994.1199