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J. Biol. Chem., Vol. 275, Issue 45, 35486-35490, November 10, 2000
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From the Department of Molecular Medicine, Chiba University
Graduate School of Medicine, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan
The Na+-driven
Cl
The Na+-driven
Cl
/HCO3
Exchanger
CLONING, TISSUE DISTRIBUTION, AND FUNCTIONAL
CHARACTERIZATION*,
,
/HCO3
exchanger is an important regulator of intracellular pH in various
cells, but its molecular basis has not been determined. We show here
the primary structure, tissue distribution, and functional
characterization of Na+-driven
chloride/bicarbonate exchanger
(designated NCBE) cloned from the insulin-secreting cell line MIN6
cDNA library. The NCBE protein consists of 1088 amino acids having
74, 72, and 55% amino acid identity to the human skeletal muscle, rat
smooth muscle, and human kidney sodium bicarbonate cotransporter,
respectively. The protein has 10 putative membrane-spanning regions.
NCBE mRNA is expressed at high levels in the brain and the mouse
insulinoma cell line MIN6 and at low levels in the pituitary, testis,
kidney, and ileum. Functional analyses of the NCBE protein expressed in Xenopus laevis oocytes and HEK293 cells demonstrate that it
transports extracellular Na+ and
HCO3 into cells in exchange
for intracellular Cl and H+, thus raising the
intracellular pH. Thus, we conclude that NCBE is a
Na+-driven
Cl/HCO3
exchanger that regulates intracellular pH in native cells.
*
This work was supported by Grant-in-aid 10NP0201 for
Creative Basic Research from the Ministry of Education, Science, Sports and Culture and by grants from the Ministry of Health and Welfare, Japan, Novo Nordisk Pharma Ltd., Yamanouchi Foundation for Research on
Metabolic Disorders, and Suzuken Memorial Foundation.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
The on-line version of this article (available at
http://www.jbc.org) contains Fig. 1S and Table IIS.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AB033759 (for NCBE).
Supported by a Japan Society for the Promotion of Science
(JSPS) postdoctoral fellowship for foreign researchers.
§
To whom correspondence should be addressed. Tel.:
81-43-226-2188; Fax: 81-43-226-2191; E-mail:
hyano@molmed.m.chiba-u.ac.jp.
¶
Supported by a JSPS research fellowship for young scientists.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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