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J Biol Chem, Vol. 274, Issue 23, 16355-16362, June 4, 1999
From the Departments of The K+-Cl
Cloning and Characterization of KCC3 and KCC4, New Members of the
Cation-Chloride Cotransporter Gene Family
,,,**, Medicine, ** Pharmacology, and
§§ Anesthesiology, Vanderbilt University Medical
Center, Nashville, Tennessee 37232 and the ¶ Molecular Physiology
Unit, Instituto Nacional de la Nutrición Salvador Zubirán
and Instituto de Investigaciones Biomédicas UNAM, Mexico City,
Mexico
cotransporters (KCCs) belong to the gene family of electroneutral
cation-chloride cotransporters, which also includes two
bumetanide-sensitive Na+-K+-2Cl
cotransporters and a thiazide-sensitive
Na+-Cl cotransporter. We have cloned
cDNAs encoding mouse KCC3, human KCC3, and human KCC4, three new
members of this gene family. The KCC3 and KCC4 cDNAs predict
proteins of 1083 and 1150 amino acids, respectively. The KCC3 and KCC4
proteins are 65-71% identical to the previously characterized
transporters KCC1 and KCC2, with which they share a predicted membrane
topology. The four KCC proteins differ at amino acid residues within
key transmembrane domains and in the distribution of putative
phosphorylation sites within the amino- and carboxyl-terminal
cytoplasmic domains. The expression of mouse KCC3 in Xenopus
laevis oocytes reveals the expected functional characteristics of
a K+Cl cotransporter:
Cl-dependent uptake of
86Rb+ which is strongly activated by cell
swelling and weakly sensitive to furosemide. A direct functional
comparison of mouse KCC3 to rabbit KCC1 indicates that KCC3 has a much
greater volume sensitivity. The human KCC3 and KCC4 genes are located
on chromosomes 5p15 and 15q14, respectively. Although widely expressed,
KCC3 transcripts are the most abundant in heart and kidney, and KCC4 is
expressed in muscle, brain, lung, heart, and kidney. The unexpected
molecular heterogeneity of K+-Cl cotransport
has implications for the physiology and pathophysiology of a number of tissues.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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