Cl- and membrane potential dependence of amino acid transport across the rat renal brush border membrane

Abstract

The relative roles of the anion present and the membrane potential in the operation of each of the seven amino acid transport systems in the renal tubular brush border membrane were explored by manipulating transmembrane potential and chemical gradients across the membrane. The effect of various external anions with different permeabilities of the membrane and of valinomycin-generated K+ diffusion potential on Na+-coupled amino acid accumulation by rat renal brush border membrane vesicles was examined. Accumulation of all amino acids examined, except for cystine, was membrane potential dependent. The highest voltage dependence was observed for taurine (equivalent to glucose) and l-methionine. Addition of taurine uptake values obtained under each electrical gradient (inside negative) and a chemical gradient (100 mM NaCl out) condition yielded markedly lower values than under conditions where there was a combined electrochemical gradient. Cl- gradient rather than merely imposing a voltage gradient was a specific mediator of Na+-coupled transport of l-proline, taurine, l-glutamic acid, and glycine across the brush border membrane. Cl- gradient alone under Na+-equilibrated conditions could energize an overshoot of taurine accumulation by vesicles providing evidence that taurine is energetically activated by and coupled to Cl- transport. These data suggest that Na+-linked transport of most amino acids across the tubular luminal membrane is an electrogenic positive process and for proline, taurine, glutamic acid, and glycine, a Cl--requiring process. A negative intracellular potential combined with luminal chloride is required for optimal Na+-coupled transport of these amino acids across the luminal membrane of the proximal tubule. The coupling of Cl- to the transport of these osmoprotective amino acids may enhance their volume regulatory effect in kidney cells and other mammalian cells.