Early metabolic effects and mechanism of ammonium transport in yeast

Abstract

Studies were performed to define the effects and mechanism of NH 4 + transport in yeast. The following results were obtained. Glucose was a better facilitator than ethanol-H 2O 2 for ammonium transport; low concentrations of uncouplers or respiratory inhibitors could inhibit the transport with ethanol as the substrate. With glucose, respiratory inhibitors showed only small inhibitory effects, and only high concentrations of azide or trifluoromethoxy carbonylcyanide phenylhydrazone could inhibit ammonium transport. Ammonium in the free state could be concentrated approximately 200-fold by the cells. Also, the addition of ammonium produced (a) stimulation of both respiration and fermentation; (b) an increased rate of H + extrusion and an alkalinization of the interior of the cell; (c) a decrease of the membrane potential, as monitored by fluorescent cyanine; (d) an immediate decrease of the levels of ATP and an increase of ADP, which may account for the stimulation of both fermentation and respiration; and (e) an increase of the levels of inorganic phosphate. Ammonium was found to inhibit 86Rb + transport much less than K +. Also, while K + produced a competitive type of inhibition, that produced by NH 4 + was of the noncompetitive type. From the distribution ratio of ammonium and the pH gradient, an electrochemical potential gradient of around −180 mV was calculated. The results indicate that ammonium is transported in yeast by a mechanism similar to that of monovalent alkaline cations, driven by a membrane potential. The immediate metabolic effects of this cation seem to be due to an increased [H +]ATPase, to which its transport is coupled. However, the carriers seem to be different. The transport system studied in this work was that of low affinity.