Possible energization of K + accumulation into metabolizing yeast by the protonmotive force binding correction to be applied in the calculation of the yeast membrane potential from tetraphenylphosphonium distribution

Abstract

Membrane potentials of yeast cells, Saccharomyces cerevisiae, calculated from the equilibrium distribution of tetraphenylphosphonium (TPP) between cell-water and medium should be corrected for a contribution due to binding of TPP to intracellular constituents. The magnitude of this correction depends upon the way in which it is determined. In cells permeabilized by boiling, cell-binding is much higher than in cells permeabilized by repeated freezing and thawing. The binding corrections are 75 ± 1 mV and 49 ± 7 mV, respectively. The binding correction obtained from TPP distribution between deenergized cells and medium is much lower and amounts to 19 ± 9 mV. The latter value is probably more reliable. It is supposed that permeabilization of the cells by boiling or repeated freezing and thawing unmasks potential TPP binding groups in the cell. The K + accumulation into anaerobically metabolizing yeast cells can be accounted for almost quantitatively by a cotransport of protons and K + ions if the lower binding correction is applied. This means that K + accumulation into the yeast cell may be driven by the sum of the protonmotive force and the membrane potential.