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lüll The recombinant human TRPV6 channel functions as Ca2+ sensor in human embryonic kidney and rat basophilic leukemia cells Bodding M; Wissenbach U; Flockerzi VJ Biol Chem 2002[Sep]; 277 (39): 36656-64The activation mechanism of the recently cloned human transient receptor potential vanilloid type 6 (TRPV6) channel, originally termed Ca(2+) transporter-like protein and Ca(2+) transporter type 1, was investigated in whole-cell patch-clamp experiments using transiently transfected human embryonic kidney and rat basophilic leukemia cells. The TRPV6-mediated currents are highly Ca(2+)-selective, show a strong inward rectification, and reverse at positive potentials, which is similar to store-operated Ca(2+) entry in electrically nonexcitable cells. The gating of TRPV6 channels is strongly dependent on the cytosolic free Ca(2+) concentration; lowering the intracellular free Ca(2+) concentration results in Ca(2+) influx, and current amplitude correlates with the intracellular EGTA or BAPTA concentration. This is also the case for TRPV6-mediated currents in the absence of extracellular divalent cations; compared with endogenous currents in nontransfected rat basophilic leukemia cells, these TRPV6-mediated monovalent currents reveal differences in reversal potential, inward rectification, and slope at very negative potentials. Release of stored Ca(2+) by inositol 1,4,5-trisphosphate and/or the sarco/endoplasmic reticulum Ca(2+)-ATPase inhibitor thapsigargin appears not to be involved in TRPV6 channel gating in both cell lines but, in rat basophilic leukemia cells, readily activates the endogenous Ca(2+) release-activated Ca(2+) current. In conclusion, TRPV6, expressed in human embryonic kidney cells and in rat basophilic leukemia cells, functions as a Ca(2+)-sensing Ca(2+) channel independently of procedures known to deplete Ca(2+) stores.|Animals[MESH]|Blotting, Northern[MESH]|Calcium Channels/*metabolism/*physiology[MESH]|Calcium-Transporting ATPases/metabolism[MESH]|Calcium/*metabolism[MESH]|Cations[MESH]|Cell Line[MESH]|Chelating Agents/pharmacology[MESH]|Cytosol/metabolism[MESH]|DNA, Complementary/metabolism[MESH]|Egtazic Acid/*analogs & derivatives/pharmacology[MESH]|Electrophysiology[MESH]|Humans[MESH]|Inositol 1,4,5-Trisphosphate/metabolism[MESH]|Leukemia/*metabolism[MESH]|Rats[MESH]|Recombinant Proteins/chemistry/metabolism[MESH]|Sarcoplasmic Reticulum Calcium-Transporting ATPases[MESH]|TRPV Cation Channels[MESH]|Thapsigargin/pharmacology[MESH]|Time Factors[MESH]|Transfection[MESH]|Tumor Cells, Cultured[MESH] |