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The electrogenicity of the rat sodium–bicarbonate cotransporter NBCe1 requires interactions among transmembrane segments of the transporter
Author(s) -
Choi Inyeong,
Soo Yang Han,
Boron Walter F.
Publication year - 2007
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2006.114959
Subject(s) - cotransporter , chemistry , intracellular ph , membrane potential , reversal potential , biophysics , reabsorption , extracellular , voltage clamp , symporter , transporter , transmembrane domain , intracellular , sodium , biochemistry , patch clamp , membrane , biology , receptor , organic chemistry , gene
The electrogenic Na + –HCO 3 − cotransporter (NBCe1) plays a central role in intracellular pH (pH i ) regulation as well as HCO 3 − secretion by pancreatic ducts and HCO 3 − 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% CO 2 –26 m m HCO 3 − (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 HCO 3 − ‐free solution) and a slope conductance of 3.0 μS (compared with 12.5 μS 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 μS), the cytoplasmic C‐terminal domain (Ct; −123.8 mV/9.7 μS) or Nt + L + Ct (−120.9 mV/3.7 μS). 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 CO 2 –HCO 3 − . pH i recovered from an acid load, but without the negative shift of V m that is characteristic of electrogenic Na + –HCO 3 − 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.