Analysis of the mKir2.1 channel activity in potassium influx defective Saccharomyces cerevisiae strains determined as changes in growth characteristics

Abstract

Potassium uptake defective Saccharomyces cerevisiae strains (Δtrk1,2 and Δtrk1,2 Δtok1) were used for the phenotypic analysis of the mouse inward rectifying Kir2.1 channel by growth analysis. Functional expression of both, multi‐copy plasmid and chromosomally expressed GFP‐mKir2.1 fusion constructs complemented the potassium uptake deficient phenotype in a pH_out dependent manner. Upon application of Hygromycin B to chromosomally mKir2.1 expressing cells, significantly lower toxin sensitivity (EC₅₀ 15.4 μM) compared to Δtrk1,2 Δtok1 cells (EC₅₀ 2.6 μM) was observed. Growth determination of mKir2.1 expressing strains upon application of Ag⁺, Cs⁺ and Ba²⁺ as known blockers of mKir2.1 channels revealed significantly decreased channel function. Cells with mKir2.1 were about double sensitive to AgNO₃, 350‐fold more sensitive to CsCl and 1500‐fold more sensitive to BaCl₂ in comparison to the respective controls indicating functional expression and correct pharmacology.