Pitfalls in the measurement of membrane potential in yeast cells using tetraphenylphosphonium

Abstract

The uptake of the lipophilic cation tetraphenylphosphonium (Ph 4P +) by Saccharomyces cerevisiae was measured using yeast grown on glucose and harvested either at the logarithmic or at the stationary phase of growth. When yeast was collected at the stationary phase, Ph 4P + uptake proceeded steadily during several hours until an equilibrium was reached. When yeast was collected in the logarithmic phase of growth, a biphasic uptake was observed. The second phase of uptake began when the glucose of the incubation medium had been exhausted. From experiments in the presence of cycloheximide or chloramphenicol it is concluded that the second phase of Ph 4P + uptake is dependent on the synthesis of some protein(s) repressed by glucose but unrelated with the existence of functional mitochondria. The addition of compounds which collapse the membrane potential provokes an efflux from the yeast cells of the Ph 4P + accumulated both during the first phase and the second phase of uptake. It is concluded that accumulation of Ph 4P + in yeast cells is a complex process and that Ph 4P + cannot be used to give a quantitative measure of the yeast plasma membrane potential.