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cotransporter from rat kidney
1 Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520; and 2 Renal Division, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115
We have recently
cloned the renal electrogenic
Na+-bicarbonate cotransporter of
the salamander Ambystoma tigrinum
(aNBC) (M. F. Romero, M. A. Hediger, E. L. Boulpaep, and W. F. Boron. FASEB J. 10: 89, 1996; and
Nature 387: 409-413, 1997). Here
we report the cloning of a mammalian homolog of aNBC, named rNBC for
rat Na+-bicarbonate cotransporter.
NBC constitutes the major route for 
reabsorption and assists in
Na+ reabsorption across the
basolateral membrane of the renal proximal tubule (PT). We used aNBC as
a probe to screen a rat kidney cortex cDNA library in 
gt10 and
identified several clones. Each has an initiator Met and a large
open-reading frame followed by a 3'-untranslated region of ~500
bp. The 7.5-kb mRNA for rNBC is present in kidney, liver, lung, brain,
and heart. In situ hybridization with the rNBC probe in the rat kidney
revealed staining in the S2 segment of PT. rNBC encodes a
protein of 1,035 amino acids, with a predicted molecular mass of 116 kDa. Its deduced amino acid sequence is 86% identical to that of aNBC.
Comparison of both the aNBC and rNBC sequences to the GenBank database
reveals a low level of amino acid identity (~30%) to the AE family
of Cl
/
exchangers. Injection of rNBC cRNA into Xenopus oocytes leads to expression of
an electrogenic
Na+-
cotransporter that is qualitatively similar to that of aNBC but at a
much lower level. Placement of the rNBC cDNA into the context of a
Xenopus expression vector produces a
substantial increase in rNBC expression. Addition of 1.5%
CO2/10 mM
elicits a hyperpolarization of
>50 mV and a rapid decrease of intracellular pH
(pHi), followed by an increase
in pHi. Subsequent
removal of Na+ in the presence of
CO2/
causes a depolarization of >50 mV and a concomitant decrease of
pHi. Thus rNBC is in the same
newly identified family of
Na+-linked
transporters as is aNBC.
rat sodium-bicarbonate cotransport; homology cloning; Xenopus oocyte expression; intracellular pH; sodium transport; bicarbonate transport
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