Solute transport across the tonoplast of barley mesophyll vacuoles: Mg²⁺ determines the specificity, and ATP and lipophilic amino acids the activity of the amino acid carrier

Abstract

After Stimulation with ATP and in the absence of divalent cations, isolated barley mesophyll vacuoles exhibited massive solute fluxes across the tonoplast, measured either as efflux of endogenous solutes or as uptake of radioactive-labeled compounds. Transported solutes were ions (particularly K⁺, NO ⁻₃ , Cl⁻) and amino acids (for example, ala, arg, asp, gln, leu, met). Addition of Mg²⁺in excess of added ATP inhibited fluxes of inorganic ions and of positively charged amino acids, but not, or to a smaller extent, those of neutral amino acids. Thus, Mg²⁺ increased the specificity of the carrier for amino acids such as alanine and glutamine. All ATP-stimulated transport processes were sensitive towards inhibition by lipophilic amino acids, for example by leucine and phenylalanine. After stimulation with sulfhydryl reagents, the inhibitory properties of Mg²⁺ and lipophilic amino acids were lost. These data concur with the hypothesis of a single transporter which exhibits a channel-like structure with a low degree of substrate selectivity in the absence of Mg²⁺, and which functions as a neutral amino acid carrier in the presence of Mg²⁺.