Characteristics of renal Rhbg as an NH4(+) transporter.

ABSTRACT

Rhbg is one of two recently cloned nonerythroid glycoproteins belonging to the Rh antigen family. Rhbg is expressed in basolateral membranes of intercalated cells of the kidney cortical collecting duct and some other cell types of the distal nephron and may function as NH(4)(+) transporters. The aim of this study was to characterize the role of Rhbg in transporting NH(4)(+). To do so, we expressed Rhbg in Xenopus laevis oocytes. Two-electrode voltage-clamp and H(+)-selective microlectrodes were used to measure NH(4)(+) currents, current-voltage plots, and intracellular pH (pH(i)). In oocytes expressing Rhbg, 5 mM NH(4)(+) induced an inward current of 93 +/- 7.7 nA (n = 20) that was significantly larger than that in control oocytes of -29 +/- 7.1 nA (P < 0.005). Whole cell conductance, at all tested potentials (-60 to +60 mV), was significantly more in oocytes expressing Rhbg compared with H(2)O-injected oocytes. In Rhbg oocytes, 5 mM NH(4)(+) depolarized the oocyte by 28 +/- 3.6 mV and decreased pH(i) by 0.30 +/- 0.04 at a rate of -20 +/- 2.5 x 10(-4) pH/s. In control oocytes, 5 mM NH(4)(+) depolarized V(m) by only 20 +/- 5.8 mV and pH(i) decreased by 0.07 +/- 0.01 at a rate of -2.7 +/- 0.6 x 10(-4) pH/s. Raising bath [NH(4)(+)] in increments from 1 to 20 mM elicited a proportionally larger decrease in pH(i) (DeltapH(i)), larger depolarization (DeltaV(m)), and a faster rate of pH(i) decrease. Bathing Rhbg oocytes in 20 mM NH(4)(+) induced an inward current of 140 +/- 7 nA that was not significantly different from 178 +/- 23 nA induced in H(2)O-injected (control) oocytes. The rate of pH(i) decrease induced by increasing external [NH(4)(+)] was significantly faster in Rhbg than in H(2)O-injected oocytes at all external NH(4)(+) concentrations. In oocytes expressing Rhbg, net NH(4)(+) influx (estimated from NH(4)(+)-induced H(+) influx) as a function of external [NH(4)(+)] saturated at higher [NH(4)(+)] with a V(max) of approximately 30.8 and an apparent K(m) of 2.3 mM (R(2) = 0.99). These data strongly suggest that Rhbg is a specific electrogenic transporter of NH(4)(+).