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Application of a membrane fusion assay for rapid drug screening
S S Vogel1, S Beushausen, D S Lester
1Laboratory of Theoretical and Physical Biology, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA.
Purpose:
The purpose of this study is to develop an in vitro assay for screening drug and their effects on membrane fusion and lysis of intracellular organelles.
Methods:
A 96-well microtiter-dish turbidimetric assay using membrane components of the eggs of sea urchins, a marine invertebrate, was applied to monitor granule fusion and/or lysis.
Results:
Of 18 drugs screened, 16 had no effect. One antineoplastic drug, tamoxifen, disrupted intracellular membranes in a calcium independent manner. Taxol, another antineoplastic drug, specifically inhibited calcium triggered exocytosis.
Conclusions:
This assay is inexpensive, simple, rapid, and does not require the sacrifice of animal life. It has the potential to identify drugs that are membrane active, as well as those which specifically perturb events involved in the secretion process.
Insights
A new assay screens drugs for effects on cell membranes using sea urchin eggs. Tamoxifen disrupted membranes, while Taxol inhibited secretion, demonstrating the assay's potential for identifying membrane-active drugs.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Membrane fusion and organelle lysis are critical cellular processes.
- Identifying drugs that modulate these processes is essential for therapeutic development.
Purpose of the Study:
- To develop a novel in vitro assay for screening drug effects on membrane fusion and organelle lysis.
- To assess the assay's utility in identifying membrane-active compounds.
Main Methods:
- A 96-well microtiter-dish turbidimetric assay was employed.
- Membrane components from sea urchin eggs were utilized as the biological system.
- The assay monitored granule fusion and/or lysis.
Main Results:
- 16 out of 18 screened drugs showed no effect on membrane integrity or fusion.
- Tamoxifen, an antineoplastic drug, disrupted intracellular membranes independently of calcium.
- Taxol, another antineoplastic drug, specifically inhibited calcium-triggered exocytosis.
Conclusions:
- The developed assay is cost-effective, simple, rapid, and ethically sound, avoiding animal sacrifice.
- This assay can effectively identify drugs that are membrane-active.
- The assay has the potential to identify drugs that specifically interfere with secretion processes.