REGULATION OF CELL PH BY K+/H+ ANTIPORT IN RENAL EPITHELIAL-CELLS

Acid-loaded opossum kidney (OK) cells secrete H+ by Na+/H+ exchange and by a Na+- and HCO3 -independent pathway that has not been fully characterized. We studied the Na+-independent component by measuring H+ flux using the pH-sensitive trapped indicator 2',7'-bis(2-carboxyethyl)-5(6)carboxyfluorescein. Two Na+-independent H+-transport systems were identified in acid-loaded cells perfused with HCO3- free buffers. The minor component appears to be a conductive pathway for H+, over 90% inhibitable by 5 mM barium. The major component is stimulated by extracellular K+ and was fully active in the presence of barium, amiloride, ouabain, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, and bumetanide and in the absence of Cl-. Ammonium inhibited the H+ flux by 72% at 50 mM, and the H+ flux could be accelerated two- to threefold by limited proteolysis of intact cells using kallikrein or papain. In cells pretreated with barium, the K+-induced H+ flux caused no change of bis-oxonol fluorescence, suggesting an electroneutral pathway. The H+ flux was a saturable function of extracellular K+ (Michaelis constant 55 mM), and flux reversed when the K+ gradient was reversed. Similarly, the H+ flux was a linear function of the H+ gradient and reversed when the H+ gradient reversed Evidence for ongoing K+-induced H+ flux was also found in nonacidified cells. First, changing perfusate K+ from 5 to 50 mM alkalinized baseline cell pH, an effect not reproduced by barium despite an equivalent depolarizing effect. Second, increasing perfusate K+ from 5 to 50 mM completely eliminated the acidification produced by 1 mM amiloride. We conclude that the OK cell expresses two Na+-independent acid-base transport systems. One is a barium-sensitive electrogenic H+ conductance and the other functions as an electroneutral K+/H+ antiporter. The antiporter is capable of H+ extrusion from acid-loaded cells but in normal cells functions in the reverse direction, as an H+ loader. The K+/H+ antiporter appears to be one of the major systems regulation cell pH in these cells, balancing the H+ efflux mediated by Na+/H+ exchange.