The electrogenicity of the rat sodium-bicarbonate cotransporter NBCe1 requires interactions among transmembrane segments of the transporter

The electrogenic Na+-HCO3- cotransporter (NBCe1) plays a central role in intracellular pH (pH(i)) regulation as well as HCO3- secretion by pancreatic ducts and HCO3- reabsorption by renal proximal tubules. To understand the structural requirements for the electrogenicity of NBCe1, we constructed chimeras of NBCe1-A and the electroneutral NBCn1-B, and used two-electrode voltage clamp to measure electrogenic transporter current in Xenopus oocytes exposed to 5% CO2-26 mM HCO3-(pH 7.40). The chimera consisting of NBCe1-A (i.e. NBCe1-A 'background') with the cytoplasmic N-terminal domain (Nt) of NBCn1-B had a reversal potential of -156.3 mV (compared with a membrane potential V-m of -43.1 mV in a HCO3--free solution) and a slope conductance of 3.0 mu S (compared with 12.5 mu S for NBCe1-A). Also electrogenic were chimeras with an NBCe1-A background but with NBCn1-B contributing the extracellular loop (L) between transmembrane segment (TM) 5 and 6 (-140.9 mV/11.1 mu S), the cytoplasmic C-terminal domain (Ct; -123.8 mV/9.7 mu S) or Nt + L + Ct (-120.9 mV/3.7 mu S). Reciprocal chimeras (with an NBCn1 background but with NBCe1 contributing Nt, L, Ct or Nt + L + Ct) produced no measurable electrogenic transporter currents in the presence of CO2-HCO3-. pH(i) recovered from an acid load, but without the negative shift of V-m that is characteristic of electrogenic Na+-HCO3- cotransporters. Thus, these chimeras were electroneutral, as were two others consisting of NBCe1(Nt-L)/NBCn1(TM6-Ct) and NBCn1(Nt-L)/NBCe1(TM6-Ct). We propose that the electrogenicity of NBCe1 requires interactions between TM1-5 and TM6-13.