Ammonium transport by the colonic H(+)-K(+)-ATPase expressed in Xenopus oocytes

Functional expression of the rat colonic H(+)-K(+)-ATPase was obtained by coexpressing its catalytic alpha-subunit and the beta(1)-subunit of the Na(+)-K(+)-ATPase in Xenopus laevis oocytes. We observed that, in oocytes expressing the rat colonic H(+)-K(+)-ATPase but not in control oocytes (expressing beta(1) alone), NH(4)Cl induced a decrease in (86)Rb uptake and the initial rate of intracellular acidification induced by extracellular NH(4)Cl was enhanced, consistent with NH(+)(4) influx via the colonic H(+)-K(+)-ATPase. In the absence of extracellular K(+), only oocytes expressing the colonic H(+)-K(+)-ATPase were able to acidify an extracellular medium supplemented with NH(4)Cl. In the absence of extracellular K(+) and in the presence of extracellular NH(+)(4), intracellular Na(+) activity in oocytes expressing the colonic H(+)-K(+)-ATPase was lower than that in control oocytes. A kinetic analysis of (86)Rb uptake suggests that NH(+)(4) acts as a competitive inhibitor of the pump. Taken together, these results are consistent with NH(+)(4) competition for K(+) on the external site of the colonic H(+)-K(+)-ATPase and with NH(+)(4) transport mediated by this pump.