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      Cloning and functional expression of a rat kidney extracellular calcium/polyvalent cation-sensing receptor.

      Proceedings of the National Academy of Sciences of the United States of America

      Xenopus laevis, Amino Acid Sequence, physiology, chemistry, biosynthesis, Receptors, Cell Surface, Receptors, Calcium-Sensing, Rats, Sprague-Dawley, Rats, analysis, RNA, Messenger, RNA, Complementary, Protein Structure, Secondary, Organ Specificity, Oocytes, Molecular Sequence Data, Male, metabolism, Kidney, In Situ Hybridization, Gene Library, Gene Expression, Female, Cloning, Molecular, Chlorides, Blotting, Northern, Base Sequence, Animals

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          The maintenance of a stable extracellular concentration of ionized calcium depends on the integrated function of a number of specialized cells (e.g., parathyroid and certain kidney epithelial cells). We recently identified another G protein-coupled receptor (BoPCaRI) from bovine parathyroid that responds to changes in extracellular Ca2+ within the millimolar range and provides a key mechanism for regulating the secretion of parathyroid hormone. Using an homology-based strategy, we now report the isolation of a cDNA encoding an extracellular Ca2+/polyvalent cation-sensing receptor (RaKCaR) from rat kidney. The predicted RaKCaR protein shares 92% identity with BoPCaR1 receptor and features a seven membrane-spanning domain, characteristic of the G protein-coupled receptors, which is preceded by a large hydrophilic extracellular NH2 terminus believed to be involved in cation binding. RaKCaR cRNA-injected Xenopus oocytes responded to extracellular Ca2+, Mg2+, Gd3+, and neomycin with characteristic activation of inositol phospholipid-dependent, intracellular Ca(2+)-induced Cl- currents. In rat kidney, Northern analysis revealed RaKCaR transcripts of 4 and 7 kb, and in situ hybridization showed localization primarily in outer medulla and cortical medullary rays. Our results provide important insights into the molecular structure of an extracellular Ca2+/polyvalent cation-sensing receptor in rat kidney and provide another basis on which to understand the role of extracellular divalent cations in regulating kidney function in mineral metabolism.

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