pH sensitivity of ammonium transport by Rhbg.
Rhbg is a membrane glycoprotein that is involved in NH(3)/NH(4)(+) transport. Several models have been proposed to describe Rhbg, including an electroneutral NH(4)(+)/H(+) exchanger, a uniporter, an NH(4)(+) 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 NH(4)(+)-induced [and methyl ammonium (MA(+))] currents and changes in intracellular pH (pH(i)), respectively. In oocytes expressing Rhbg, 5 mM NH(4)Cl (NH(3)/NH(4)(+)) 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 NH(4)(+)-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 NH(4)(+)-induced current and pH(i) decrease. In oocytes expressing Rhbg, 5 mM methyl amine hydrochloride (MA/MA(+)), often used as an NH(4)Cl 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 NH(4)(+) and MA/MA(+) by Rhbg is pH sensitive; 2) electrogenic NH(4)(+) 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.
Nakhoul
Nazih L
NL
Section of Nephrology, Department of Medicine, Tulane Hypertension and Renal Center of Excellence, Tulane University School of Medicine, New Orleans, Louisiana 70112, USA. nakhoul@tulane.edu
Abdulnour-Nakhoul
Solange M
SM
Schmidt
Eric
E
Doetjes
Rienk
R
Rabon
Edd
E
Hamm
L Lee
LL
D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008564:Membrane Potentials / Q000187:drug effects; D026901:Membrane Transport Proteins / Q000378:metabolism*; D008744:Methylamines / Q000378:metabolism; D051379:Mice; D000644:Quaternary Ammonium Compounds / Q000378:metabolism*
D002087:Butyrates; D006023:Glycoproteins; D026901:Membrane Transport Proteins; D008744:Methylamines; D000644:Quaternary Ammonium Compounds; C486033:RhBG protein, mouse
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