pH sensitivity of ammonium transport by Rhbg

Nakhoul NL, Abdulnour-Nakhoul SM, Schmidt E, Doetjes R, Rabon E, Hamm LL. pH sensitivity of ammonium transport by Rhbg. Am J Physiol Cell Physiol 299: C1386-C1397, 2010. First published September 1, 2010; doi:10.1152/ajpcell.00211.2010.-Rhbg is a membrane glycoprotein that is involved in NH3/NH4+ transport. Several models have been proposed to describe Rhbg, including an electroneutral NH4+/H+ exchanger, a uniporter, an NH4+ channel, or even a gas channel. In this study, we characterized the pH sensitivity of Rhbg expressed in Xenopus oocytes. We used two-electrode voltage clamp and ion-selective microelectrodes to measure NH4+-induced [and methyl ammonium (MA(+))] currents and changes in intracellular pH (pH(i)), respectively. In oocytes expressing Rhbg, 5 mM NH4Cl (NH3/NH4+) at extracellular pH (pH(o)) of 7.5 induced an inward current, decreased pH(i), and depolarized the cell. Raising pH(o) to 8.2 significantly enhanced the NH4+-induced current and pH(i) changes, whereas decreasing bath pH to 6.5 inhibited these changes. Lowering pH(i) (decreased by butyrate) also inhibited the NH4+-induced current and pH(i) decrease. In oocytes expressing Rhbg, 5 mM methyl amine hydrochloride (MA/MA(+)), often used as an NH4Cl substitute, induced an inward current, a pH(i) increase (not a decrease), and depolarization of the cell. Exposing the oocyte to MA/MA(+) at alkaline bath pH (8.2) enhanced the MA(+)-induced current, whereas lowering bath pH to 6.5 inhibited the MA(+) current completely. Exposing the oocyte to MA/MA(+) at low pH(i) abolished the MA(+)-induced current and depolarization; however, pH(i) still increased. These data indicate that 1) transport of NH4+ and MA/MA(+) by Rhbg is pH sensitive; 2) electrogenic NH4+ and MA(+) transport are stimulated by alkaline pH(o) but inhibited by acidic pH(i) or pH(o); and 3) electroneutral transport of MA by Rhbg is likely but is less sensitive to pH changes.