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J. Biol. Chem., Vol. 275, Issue 48, 37765-37773, December 1, 2000
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From the Renal Division and Membrane Biology Program, Department of
Medicine, Brigham and Women's Hospital and Harvard Medical School,
Boston, Massachusetts 02115
CLC5 is an intracellular chloride channel of
unknown function, expressed in the renal proximal tubule. The
subcellular localization and function of CLC5 were investigated in the
LLC-PK1 porcine proximal tubule cell line. We cloned a cDNA for the
porcine CLC5 ortholog (pCLC5) that is predicted to encode an 83-kDa
protein with 97% amino acid sequence identity to rat and human CLC5.
By immunofluorescence, pCLC5 was localized to early endosomes of the
apical membrane fluid-phase endocytotic pathway and to the Golgi
complex. Xenopus oocytes injected with pCLC5 cRNA exhibited outwardly rectifying whole cell currents with a relative conductance profile (nitrate The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF274055.
Molecular Cloning and Characterization of an Intracellular
Chloride Channel in the Proximal Tubule Cell Line, LLC-PK1*
Cl
Br > I > acetate > gluconate) different from that of control oocytes. Acidification of the
extracellular medium reversibly inhibited this outward current with a
pKa of 6.0 and a Hill coefficient of 1. Overexpression of CLC5 in LLC-PK1 cells resulted in morphological changes, including loss of cell-cell contacts and the appearance of
multiple prominent vesicles. These findings are consistent with a
potential role for CLC5 in the acidification of membrane compartments
of both the endocytic and the exocytic pathway and suggest that its
function may be important for normal intercellular adhesion and
vesicular trafficking.
*
This work was supported by a Carl W. Gottschalk Research
Scholar award from the American Society of Nephrology (to A. Y.).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.
To whom correspondence should be addressed: Renal Division,
Brigham and Women's Hospital, 77 Avenue Louis Pasteur, Boston, MA
02115. Tel.: 617-525-5835; Fax: 617-525-5836; E-mail:
ayu@rics.bwh.harvard.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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