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J. Biol. Chem., Vol. 275, Issue 36, 28186-28194, September 8, 2000
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From the Active absorption of calcium from the
intestine and reabsorption of calcium from the kidney are major
determinants of whole body calcium homeostasis. Two recently
cloned proteins, CaT1 and ECaC, have been postulated to mediate apical
calcium uptake by rat intestine and rabbit kidney, respectively.
By screening a rat kidney cortex library with a CaT1 probe, we isolated
a cDNA encoding a protein (CaT2) with 84.2 and 73.4% amino acid
identities to ECaC and CaT1, respectively. Unlike ECaC, CaT2 is
kidney-specific in the rat and was not detected in intestine, brain,
adrenal gland, heart, skeletal muscle, liver, lung, spleen, thymus, and
testis by Northern analysis or reverse transcription polymerase chain reaction. The expression pattern of CaT2 in kidney was similar to that of calbindin D28K and the sodium calcium
exchanger 1, NCX1, by in situ hybridization of adjacent
sections. Furthermore, the mRNAs for CaT2 and calbindin
D28K were colocalized in the same cells. CaT2 mediated
saturable calcium uptake with a Michaelis constant
(Km) of 0.66 mM when expressed in
Xenopus laevis oocytes. Under voltage clamp condition, CaT2
promoted inward currents in X. laevis oocytes upon external
application of Ca2+. Sr2+ and Ba2+
but not Mg2+ also evoked inward currents in CaT2-expressing
oocytes. Similar to the alkaline earth metal ions, application of
Cd2+ elicited inward current in CaT2-expressing oocytes
with a Km of 1.3 mM. Cd2+,
however, also potently inhibited CaT2-mediated Ca2+
uptake with an IC50 of 5.4 µM.
Ca2+ evoked currents were reduced at low pH and increased
at high pH and were only slightly affected by the L-type
voltage-dependent calcium channel antagonists, nifedipine,
verapamil, diltiazem, and the agonist, Bay K 8644, even at relatively
high concentrations. In conclusion, CaT2 may participate in calcium
entry into the cells of the distal convoluted tubule and connecting
segment of the nephron, where active reabsorption of calcium takes
place via the transcellular route. The high sensitivity of CaT2 to
Cd2+ also provides a potential explanation for
Cd2+-induced hypercalciuria and resultant renal stone formation.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AF209196.
A Rat Kidney-specific Calcium Transporter in the Distal
Nephron*
§,§¶,§,
,, and§**
Membrane Biology Program and
§ Renal and Endocrine-Hypertension Divisions,
Department of Medicine, Brigham and Women's Hospital and Harvard
Medical School, Boston, Massachusetts 02115
*
This work was supported by a Brigham and Women's Hospital
dual-mentored fellowship (to J.-B. P.) (mentors: M. A. H. and
E. M. B.); National Institutes of Health Grants DK41415, DK48330, and
DK52005 (to M. A. H. and E. M. B.); and a grant from the St. Giles Foundation (to E. M. B. and P. M. V.). Funding was also supplied by the National Dairy Council (to E. M. B.).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.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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