Relative CO₂/NH₃ permeabilities of human RhAG, RhBG and RhCG.
Mammalian glycosylated rhesus (Rh) proteins include the erythroid RhAG and the nonerythroid RhBG and RhCG. RhBG and RhCG are expressed in multiple tissues, including hepatocytes and the collecting duct (CD) of the kidney. Here, we expressed human RhAG, RhBG and RhCG in Xenopus oocytes (vs. H2O-injected control oocytes) and used microelectrodes to monitor the maximum transient change in surface pH (DpHS) caused by exposing the same oocyte to 5 % CO₂/33 mM HCO₃⁻ (an increase) or 0.5 mM NH₃/NH₄⁺ (a decrease). Subtracting the respective values for day-matched, H₂O-injected control oocytes yielded channel-specific values (*). (ΔpH*(S))(CO₂) and (-ΔpH*(S))(NH₃) were each significantly >0 for all channels, indicating that RhBG and RhCG--like RhAG--can carry CO₂ and NH₃. We also investigated the role of a conserved aspartate residue, which was reported to inhibit NH₃ transport. However, surface biotinylation experiments indicate the mutants RhBG(D178N) and RhCG(D177N) have at most a very low abundance in the oocyte plasma membrane. We demonstrate for the first time that RhBG and RhCG--like RhAG--have significant CO₂ permeability, and we confirm that RhAG, RhBG and RhCG all have significant NH₃ permeability. However, as evidenced by (ΔpH*(S))(CO₂)/ (-ΔpH*(S))(NH₃) values, we could not distinguish among the CO₂/ NH₃ permeability ratios for RhAG, RhBG and RhCG. Finally, we propose a mechanism whereby RhBG and RhCG contribute to acid secretion in the CD by enhancing the transport of not only NH₃ but also CO₂ across the membranes of CD cells.
R Ryan
RR
Geyer
Mark D
MD
Parker
Ashley M
AM
Toye
Walter F
WF
Boron
Raif
R
Musa-Aziz
D000595:Amino Acid Sequence; D000641:Ammonia / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D001798:Blood Proteins / Q000737:chemistry / Q000378:metabolism*; D002245:Carbon Dioxide / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D015870:Gene Expression; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D008564:Membrane Potentials; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000378:metabolism*; D008845:Microinjections; D008954:Models, Biological; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D010539:Permeability; D016415:Sequence Alignment; D014981:Xenopus
D001798:Blood Proteins; D027682:Cation Transport Proteins; D006023:Glycoproteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; C498523:RHAG protein, human; C422995:RHBG protein, human; C413213:RHCG protein, human; D002245:Carbon Dioxide; D000641:Ammonia