SLC4A11 function: evidence for H⁺(OH^-) and NH₃-H⁺ transport

Whether SLC4A11 transports ammonia and its potential mode of ammonia transport ($$, NH₃, or NH₃-2H⁺ transport have been proposed) are controversial. In the absence of ammonia, whether SLC4A11 mediates significant conductive H⁺(OH^-) transport is also controversial. The present study was performed to determine the mechanism of human SLC4A11 ammonia transport and whether the transporter mediates conductive H⁺(OH^-) transport in the absence of ammonia. We quantitated H⁺ flux by monitoring changes in intracellular pH (pH_i) and measured whole cell currents in patch-clamp studies of HEK293 cells expressing the transporter in the absence and presence of NH₄Cl. Our results demonstrate that SLC4A11 mediated conductive H⁺(OH^-) transport that was stimulated by raising the extracellular pH (pH_e). Ammonia-induced HEK293 whole cell currents were also stimulated by an increase in pH_e. In studies using increasing NH₄Cl concentrations with equal $$ extracellular and intracellular concentrations, the shift in the reversal potential (E_rev) due to the addition of ammonia was compatible with NH₃-H⁺ transport competing with H⁺(OH^-) rather than NH₃-nH⁺ (n ≥ 2) transport. The increase in equivalent H⁺(OH^-) flux observed in the presence of a transcellular H⁺ gradient was also compatible with SLC4A11-mediated NH₃-H⁺ flux. The NH₃ versus E_rev data fit a theoretical model suggesting that NH₃-H⁺ and H⁺(OH^-) competitively interact with the transporter. Studies of mutant SLC4A11 constructs in the putative SLC4A11 ion coordination site showed that both H⁺(OH^-) transport and ammonia-induced whole cell currents were blocked suggesting that the H⁺(OH^-) and NH₃-H⁺ transport processes share common features involving the SLC4A11 transport mechanism.