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Finding new potential antagonists of potassium channels is a continuing task. TASK potassium channels operate over a large physiological range of membrane voltages, why they are thought to contribute to the excitability and resting potential of mammalian membrane potentials. Additionally, they are regulated by extracellular stimuli like changes in pH and K+ concentrations. TASK malfunctions are associated with diseases, which makes them popular targets for the search of new antagonists. Identification of channel inhibitors can be a time-consuming and expensive project. Here, we present an easy-to-use and inexpensive yeast system for the expression of the two-pore domain K+ channel TASK-3, and for the characterization of TASK-3 antagonists. The Saccharomyces cerevisiae strain BYT45 was used to express guinea pig TASK-3. The system allowed the expression and characterization of TASK-3 at variable pH values and K+ concentrations. Three known TASK-3 antagonists have been tested in the BYT45 yeast system: PK-THPP, ZnCl2 and Bupivacaine. Their inhibitory effect on TASK-3 was tested in solid and liquid media assays, and half maximal inhibitory concentrations were estimated. Although the system is less sensitive than more refined systems, the antagonistic activity could be confirmed for all three inhibitors.

fems yeast research

Yeast strain Saccharomyces cerevisiae BYT45 lacking the cation extrusion systems ENA1-5 and NHA1 is suitable for the characterization of TASK-3 potassium channel antagonists