Fluorescence method for determining the mechanism and speed of action of surface-active drugs on yeast cells

BioTechniques

Dana Gášková¹  Marie Kodedová¹  Karel Sigler²  Bernard D. Lemire³ 

[1] 1Charles University, Faculty of Mathematics and Physics, Institute of Physics, Prague, Czech Republic;2Institute of Microbiology, CR Academy of Sciences, Prague, Czech Republic;3Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada;

关键词: fluorescent dye diS-C3(3);    membrane potential;    surface-active drug;    cell permeabilization;   

DOI  :  10.2144/000113568

来源: DOAJ

【 摘 要 】

New antifungal agents are needed to treat life-threatening fungal infections, particularly with the development of resistance. Surface-active antifungals have the advantages of minimizing host toxicity and the emergence of drug resistance. We have developed a time-dependent drug exposure assay that allows us to rapidly investigate the mechanism of surface-active antifungal drug action. The assay uses a multidrug pump-deficient strain of Saccharomyces cerevisiae and the potentiometric dye 3,3′-dipropylthiacarbocyanine iodide [diS-C3(3)] and can assess whether cells are depolarized, hyperpolarized, or permeabilized by drug exposure. In this work, we investigated the mechanisms of action of five surface-active compounds: SDS, nystatin, amphotericin B, octenidine dihydrochloride, and benzalkonium chloride. The diS-C3(3) time-dependent drug exposure assay can be used to identify the mechanisms of action of a wide range of drugs. It is a fast and cost-effective method for screening drugs to determine their lowest effective concentrations.

【 授权许可】

Unknown