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(Received for publication, March 18, 1996)
From the Department of Cellular and Molecular Physiology, Yale
University School of Medicine, New Haven, Connecticut 06520 and the
§ Department of Human Physiology, University of California
School of Medicine, Davis, California 95616
We report the cloning, sequence analysis, tissue
distribution, and functional expression of the K-Cl cotransport
protein, KCC1. KCC1 was identified by searching the human expressed
sequence tag data base, based on the expectation that it would be
distantly related to the Na-K-Cl cotransporter. Rabbit KCC1 (rbKCC1)
and rat KCC1 (rtKCC1) were cloned by screening rabbit kidney and
rat brain cDNA libraries using homologous cDNA probes. Human
KCC1 (hKCC1) was obtained from I.M.A.G.E. clones and in part by reverse
transcription-polymerase chain reaction; it exhibits 97% identity with
rbKCC1. KCC1 encodes a 1085-residue polypeptide with substantial
sequence homology (24-25% identity) to the bumetanide-sensitive
Na-K-Cl cotransporter (NKCC or BSC) and the thiazide-sensitive Na-Cl
cotransporter (NCC or TSC). Hydropathy analysis of KCC1 indicates
structural homology to NKCC, including 12 transmembrane domains, a
large extracellular loop with potential N-linked
glycosylation sites, and cytoplasmic N- and C-terminal regions.
Northern blot analysis revealed a ubiquitously expressed 3.8-kilobase
transcript. Much of the genomic sequence of hKCC1 is in the data base,
and the gene has been previously localized to 16q22.1 (Larsen, F.,
Solhein, J., Kristensen, T., Kolsto, A. B., and Prydz, H. (1993)
Hum. Mol. Genet. 2, 1589-1595). Epitope-tagged rbKCC1 was
stably expressed in human embryonic kidney (HEK 293) cells, resulting
in production of a ~150-kDa glycoprotein. The initial rate of
86Rb efflux from cells expressing rbKCC1 was more than 7 times greater than efflux from control cells and was inhibited by 2 mM furosemide; 86Rb efflux was stimulated by
cell swelling. Uptake of 86Rb into rbKCC1 cells after a
15-min pretreatment with 1 mM N-ethylmaleimide
was dependent on external chloride but not on external sodium, and was
inhibited by furosemide with a Ki of ~40
µM and by bumetanide with a Ki of
~60 µM. These data demonstrate that the KCC1 cDNAs
encode a widely expressed K-Cl cotransporter with the characteristics
of the K-Cl transporter that has been characterized in red cells.
Volume 271, Number 27,
Issue of July 5, 1996
pp. 16237-16244
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
A NEW MEMBER OF THE CATION-CHLORIDE COTRANSPORTER FAMILY
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S. N. Orlov, N. C. Adragna, V. A. Adarichev, and P. Hamet Genetic and biochemical determinants of abnormal monovalent ion transport in primary hypertension Am J Physiol Cell Physiol, March 1, 1999; 276(3): C511 - C536. [Abstract] [Full Text] [PDF]
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C. M. Gillen and B. Forbush III Functional interaction of the K-Cl cotransporter (KCC1) with the Na-K-Cl cotransporter in HEK-293 cells Am J Physiol Cell Physiol, February 1, 1999; 276(2): C328 - C336. [Abstract] [Full Text] [PDF]
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H. Liapis, M. Nag, and D. M. Kaji K-Cl cotransporter expression in the human kidney Am J Physiol Cell Physiol, December 1, 1998; 275(6): C1432 - C1437. [Abstract] [Full Text] [PDF]
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E. J. Holtzman, S. Kumar, C. A. Faaland, F. Warner, P. J. Logue, S. J. Erickson, G. Ricken, J. Waldman, S. Kumar, and P. B. Dunham Cloning, characterization, and gene organization of K-Cl cotransporter from pig and human kidney and C. elegans Am J Physiol Renal Physiol, October 1, 1998; 275(4): F550 - F564. [Abstract] [Full Text] [PDF]
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 G. O. Andersen, M. Enger, G. H. Thoresen, T. Skomedal, and J.-B. Osnes alpha 1-Adrenergic activation of myocardial Na-K-2Cl cotransport involving mitogen-activated protein kinase Am J Physiol Heart Circ Physiol, August 1, 1998; 275(2): H641 - H652. [Abstract] [Full Text] [PDF]
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P. Isenring, S. C. Jacoby, and B. Forbush III The role of transmembrane domain 2 in cation transport by the Na-K-Cl cotransporter PNAS, June 9, 1998; 95(12): 7179 - 7184. [Abstract] [Full Text] [PDF]
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 K. L. Hoppe and O. L. Francone Binding and functional effects of transcription factors Sp1 and Sp3 on the proximal human lecithin:cholesterol acyltransferase promoter J. Lipid Res., May 1, 1998; 39(5): 969 - 977. [Abstract] [Full Text]
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P. Isenring, S. C. Jacoby, J. A. Payne, and B. Forbush III Comparison of Na-K-Cl Cotransporters. NKCC1, NKCC2, AND THE HEK CELL Na-K-Cl COTRANSPORTER J. Biol. Chem., May 1, 1998; 273(18): 11295 - 11301. [Abstract] [Full Text] [PDF]
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J. A. Payne Functional characterization of the neuronal-specific K-Cl cotransporter: implications for [K+]o regulation Am J Physiol Cell Physiol, November 1, 1997; 273(5): C1516 - C1525. [Abstract] [Full Text] [PDF]
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P. Isenring and B. Forbush III Ion and Bumetanide Binding by the Na-K-Cl Cotransporter. IMPORTANCE OF TRANSMEMBRANE DOMAINS J. Biol. Chem., September 26, 1997; 272(39): 24556 - 24562. [Abstract] [Full Text] [PDF]
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D. G. Perregaux, R. E. Laliberte, and C. A. Gabel Human Monocyte Interleukin-1[BETA] Posttranslational Processing. EVIDENCE OF A VOLUME-REGULATED RESPONSE J. Biol. Chem., November 22, 1996; 271(47): 29830 - 29838. [Abstract] [Full Text] [PDF]
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J. A. Payne, T. J. Stevenson, and L. F. Donaldson Molecular Characterization of a Putative K-Cl Cotransporter in Rat Brain. A NEURONAL-SPECIFIC ISOFORM J. Biol. Chem., July 5, 1996; 271(27): 16245 - 16252. [Abstract] [Full Text] [PDF]
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L. Caron, F. Rousseau, E. Gagnon, and P. Isenring Cloning and Functional Characterization of a Cation-Cl- Cotransporter-interacting Protein J. Biol. Chem., October 6, 2000; 275(41): 32027 - 32036. [Abstract] [Full Text] [PDF]
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A. Mercado, L. Song, N. Vazquez, D. B. Mount, and G. Gamba Functional Comparison of the K+-Cl- Cotransporters KCC1 and KCC4 J. Biol. Chem., September 22, 2000; 275(39): 30326 - 30334. [Abstract] [Full Text] [PDF]
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P. Sangan, S. R. Brill, S. Sangan, B. Forbush III, and H. J. Binder Basolateral K-Cl Cotransporter Regulates Colonic Potassium Absorption in Potassium Depletion J. Biol. Chem., September 29, 2000; 275(40): 30813 - 30816. [Abstract] [Full Text] [PDF]
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M. Di Fulvio, T. M. Lincoln, P. K. Lauf, and N. C. Adragna Protein Kinase G Regulates Potassium Chloride Cotransporter-3 Expression in Primary Cultures of Rat Vascular Smooth Muscle Cells J. Biol. Chem., June 8, 2001; 276(24): 21046 - 21052. [Abstract] [Full Text] [PDF]
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M. Di Fulvio, P. K. Lauf, and N. C. Adragna Nitric Oxide Signaling Pathway Regulates Potassium Chloride Cotransporter-1 mRNA Expression in Vascular Smooth Muscle Cells J. Biol. Chem., November 21, 2001; 276(48): 44534 - 44540. [Abstract] [Full Text] [PDF]
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M.-R. Shen, C.-Y. Chou, K.-F. Hsu, H.-S. Liu, P. B. Dunham, E. J. Holtzman, and J. C. Ellory The KCl cotransporter isoform KCC3 can play an important role in cell growth regulation PNAS, December 4, 2001; 98(25): 14714 - 14719. [Abstract] [Full Text] [PDF]
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B. Wyse, N. Ali, and D. H. Ellison Interaction with grp58 increases activity of the thiazide-sensitive Na-Cl cotransporter Am J Physiol Renal Physiol, March 1, 2002; 282(3): F424 - F430. [Abstract] [Full Text] [PDF]
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N. Vazquez, A. Monroy, E. Dorantes, R. A. Munoz-Clares, and G. Gamba Functional differences between flounder and rat thiazide-sensitive Na-Cl cotransporter Am J Physiol Renal Physiol, April 1, 2002; 282(4): F599 - F607. [Abstract] [Full Text] [PDF]
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