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. 1998 Aug 3;17(15):4274–4282. doi: 10.1093/emboj/17.15.4274

A novel capacitative calcium entry channel expressed in excitable cells.

S Philipp 1, J Hambrecht 1, L Braslavski 1, G Schroth 1, M Freichel 1, M Murakami 1, A Cavalié 1, V Flockerzi 1
PMCID: PMC1170761  PMID: 9687496

Abstract

In addition to voltage-gated calcium influx, capacitative calcium entry (CCE) represents a major pathway for calcium entry into the cell. Here we report the structure, expression and functional properties of a novel CCE channel, TRP5. This channel is a member of a new subfamily of mammalian homologues of the Drosophila transient receptor potential (TRP) protein, now comprising TRP5 (also CCE2) and the structurally related CCE1 (also TRP4). Like TRP4, TRP5 forms ion channels mainly permeable for Ca2+ which are not active under resting conditions but can be activated by manoeuvres known to deplete intracellular calcium stores. Accordingly, dialysis of TRP5-expressing cells with inositol-(1,4,5)-trisphosphate evokes inward rectifying currents which reversed polarity at potentials more positive than +30 mV. Ca2+ store depletion with thapsigargin induced TRP5-mediated calcium entry dependent on the concentration of extracellular calcium, as seen by dual wavelength fura-2 fluorescence ratio measurements. TRP5 transcripts are expressed almost exclusively in brain, where they are present in mitral cells of the olfactory bulb, in lateral cerebellar nuclei and, together with TRP4 transcripts, in CA1 pyramidal neurons of the hippocampus, indicating the presence of CCE channels in excitable cells and their participation in neuronal calcium homeostasis.

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Selected References

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  1. Birnbaumer L., Zhu X., Jiang M., Boulay G., Peyton M., Vannier B., Brown D., Platano D., Sadeghi H., Stefani E. On the molecular basis and regulation of cellular capacitative calcium entry: roles for Trp proteins. Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15195–15202. doi: 10.1073/pnas.93.26.15195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boulay G., Zhu X., Peyton M., Jiang M., Hurst R., Stefani E., Birnbaumer L. Cloning and expression of a novel mammalian homolog of Drosophila transient receptor potential (Trp) involved in calcium entry secondary to activation of receptors coupled by the Gq class of G protein. J Biol Chem. 1997 Nov 21;272(47):29672–29680. doi: 10.1074/jbc.272.47.29672. [DOI] [PubMed] [Google Scholar]
  3. Chang A. S., Chang S. M., Garcia R. L., Schilling W. P. Concomitant and hormonally regulated expression of trp genes in bovine aortic endothelial cells. FEBS Lett. 1997 Oct 6;415(3):335–340. doi: 10.1016/s0014-5793(97)01155-1. [DOI] [PubMed] [Google Scholar]
  4. Chevesich J., Kreuz A. J., Montell C. Requirement for the PDZ domain protein, INAD, for localization of the TRP store-operated channel to a signaling complex. Neuron. 1997 Jan;18(1):95–105. doi: 10.1016/s0896-6273(01)80049-0. [DOI] [PubMed] [Google Scholar]
  5. Freichel M., Wissenbach U., Philipp S., Flockerzi V. Alternative splicing and tissue specific expression of the 5' truncated bCCE 1 variant bCCE 1delta514. FEBS Lett. 1998 Feb 6;422(3):354–358. doi: 10.1016/s0014-5793(98)00041-6. [DOI] [PubMed] [Google Scholar]
  6. Funayama M., Goto K., Kondo H. Cloning and expression localization of cDNA for rat homolog of TRP protein, a possible store-operated calcium (Ca2+) channel. Brain Res Mol Brain Res. 1996 Dec 31;43(1-2):259–266. doi: 10.1016/s0169-328x(96)00208-2. [DOI] [PubMed] [Google Scholar]
  7. Garaschuk O., Yaari Y., Konnerth A. Release and sequestration of calcium by ryanodine-sensitive stores in rat hippocampal neurones. J Physiol. 1997 Jul 1;502(Pt 1):13–30. doi: 10.1111/j.1469-7793.1997.013bl.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. García D. E., Cavalié A., Lux H. D. Enhancement of voltage-gated Ca2+ currents induced by daily stimulation of hippocampal neurons with glutamate. J Neurosci. 1994 Feb;14(2):545–553. doi: 10.1523/JNEUROSCI.14-02-00545.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gillo B., Chorna I., Cohen H., Cook B., Manistersky I., Chorev M., Arnon A., Pollock J. A., Selinger Z., Minke B. Coexpression of Drosophila TRP and TRP-like proteins in Xenopus oocytes reconstitutes capacitative Ca2+ entry. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):14146–14151. doi: 10.1073/pnas.93.24.14146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gomez T. M., Snow D. M., Letourneau P. C. Characterization of spontaneous calcium transients in nerve growth cones and their effect on growth cone migration. Neuron. 1995 Jun;14(6):1233–1246. doi: 10.1016/0896-6273(95)90270-8. [DOI] [PubMed] [Google Scholar]
  11. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  12. Heim R., Cubitt A. B., Tsien R. Y. Improved green fluorescence. Nature. 1995 Feb 23;373(6516):663–664. doi: 10.1038/373663b0. [DOI] [PubMed] [Google Scholar]
  13. Hoth M., Penner R. Depletion of intracellular calcium stores activates a calcium current in mast cells. Nature. 1992 Jan 23;355(6358):353–356. doi: 10.1038/355353a0. [DOI] [PubMed] [Google Scholar]
  14. Hu Y., Schilling W. P. Receptor-mediated activation of recombinant Trpl expressed in Sf9 insect cells. Biochem J. 1995 Jan 15;305(Pt 2):605–611. doi: 10.1042/bj3050605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hu Y., Vaca L., Zhu X., Birnbaumer L., Kunze D. L., Schilling W. P. Appearance of a novel Ca2+ influx pathway in Sf9 insect cells following expression of the transient receptor potential-like (trpl) protein of Drosophila. Biochem Biophys Res Commun. 1994 Jun 15;201(2):1050–1056. doi: 10.1006/bbrc.1994.1808. [DOI] [PubMed] [Google Scholar]
  16. Huber A., Sander P., Gobert A., Bähner M., Hermann R., Paulsen R. The transient receptor potential protein (Trp), a putative store-operated Ca2+ channel essential for phosphoinositide-mediated photoreception, forms a signaling complex with NorpA, InaC and InaD. EMBO J. 1996 Dec 16;15(24):7036–7045. [PMC free article] [PubMed] [Google Scholar]
  17. Hurst R. S., Zhu X., Boulay G., Birnbaumer L., Stefani E. Ionic currents underlying HTRP3 mediated agonist-dependent Ca2+ influx in stably transfected HEK293 cells. FEBS Lett. 1998 Feb 6;422(3):333–338. doi: 10.1016/s0014-5793(98)00035-0. [DOI] [PubMed] [Google Scholar]
  18. Kaufman R. J., Davies M. V., Wasley L. C., Michnick D. Improved vectors for stable expression of foreign genes in mammalian cells by use of the untranslated leader sequence from EMC virus. Nucleic Acids Res. 1991 Aug 25;19(16):4485–4490. doi: 10.1093/nar/19.16.4485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kornau H. C., Seeburg P. H., Kennedy M. B. Interaction of ion channels and receptors with PDZ domain proteins. Curr Opin Neurobiol. 1997 Jun;7(3):368–373. doi: 10.1016/s0959-4388(97)80064-5. [DOI] [PubMed] [Google Scholar]
  20. Kozak M. An analysis of 5'-noncoding sequences from 699 vertebrate messenger RNAs. Nucleic Acids Res. 1987 Oct 26;15(20):8125–8148. doi: 10.1093/nar/15.20.8125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  22. Lückhoff A., Clapham D. E. Calcium channels activated by depletion of internal calcium stores in A431 cells. Biophys J. 1994 Jul;67(1):177–182. doi: 10.1016/S0006-3495(94)80467-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Montell C., Rubin G. M. Molecular characterization of the Drosophila trp locus: a putative integral membrane protein required for phototransduction. Neuron. 1989 Apr;2(4):1313–1323. doi: 10.1016/0896-6273(89)90069-x. [DOI] [PubMed] [Google Scholar]
  24. Montero M., Alvarez J., Garcia-Sancho J. Uptake of Ca2+ and refilling of intracellular Ca2+ stores in Ehrlich-ascites-tumour cells and in rat thymocytes. Biochem J. 1990 Oct 15;271(2):535–540. doi: 10.1042/bj2710535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mountford P., Zevnik B., Düwel A., Nichols J., Li M., Dani C., Robertson M., Chambers I., Smith A. Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4303–4307. doi: 10.1073/pnas.91.10.4303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Needleman S. B., Wunsch C. D. A general method applicable to the search for similarities in the amino acid sequence of two proteins. J Mol Biol. 1970 Mar;48(3):443–453. doi: 10.1016/0022-2836(70)90057-4. [DOI] [PubMed] [Google Scholar]
  27. Okada T., Shimizu S., Wakamori M., Maeda A., Kurosaki T., Takada N., Imoto K., Mori Y. Molecular cloning and functional characterization of a novel receptor-activated TRP Ca2+ channel from mouse brain. J Biol Chem. 1998 Apr 24;273(17):10279–10287. doi: 10.1074/jbc.273.17.10279. [DOI] [PubMed] [Google Scholar]
  28. Petersen C. C., Berridge M. J., Borgese M. F., Bennett D. L. Putative capacitative calcium entry channels: expression of Drosophila trp and evidence for the existence of vertebrate homologues. Biochem J. 1995 Oct 1;311(Pt 1):41–44. doi: 10.1042/bj3110041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Philipp S., Cavalié A., Freichel M., Wissenbach U., Zimmer S., Trost C., Marquart A., Murakami M., Flockerzi V. A mammalian capacitative calcium entry channel homologous to Drosophila TRP and TRPL. EMBO J. 1996 Nov 15;15(22):6166–6171. [PMC free article] [PubMed] [Google Scholar]
  30. Philipp S., Flockerzi V. Molecular characterization of a novel human PDZ domain protein with homology to INAD from Drosophila melanogaster. FEBS Lett. 1997 Aug 18;413(2):243–248. doi: 10.1016/s0014-5793(97)00877-6. [DOI] [PubMed] [Google Scholar]
  31. Phillips A. M., Bull A., Kelly L. E. Identification of a Drosophila gene encoding a calmodulin-binding protein with homology to the trp phototransduction gene. Neuron. 1992 Apr;8(4):631–642. doi: 10.1016/0896-6273(92)90085-r. [DOI] [PubMed] [Google Scholar]
  32. Prasher D. C., Eckenrode V. K., Ward W. W., Prendergast F. G., Cormier M. J. Primary structure of the Aequorea victoria green-fluorescent protein. Gene. 1992 Feb 15;111(2):229–233. doi: 10.1016/0378-1119(92)90691-h. [DOI] [PubMed] [Google Scholar]
  33. Preuss K. D., Nöller J. K., Krause E., Göbel A., Schulz I. Expression and characterization of a trpl homolog from rat. Biochem Biophys Res Commun. 1997 Nov 7;240(1):167–172. doi: 10.1006/bbrc.1997.7528. [DOI] [PubMed] [Google Scholar]
  34. Putney J. W., Jr A model for receptor-regulated calcium entry. Cell Calcium. 1986 Feb;7(1):1–12. doi: 10.1016/0143-4160(86)90026-6. [DOI] [PubMed] [Google Scholar]
  35. Saitou N., Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987 Jul;4(4):406–425. doi: 10.1093/oxfordjournals.molbev.a040454. [DOI] [PubMed] [Google Scholar]
  36. Sakura H., Ashcroft F. M. Identification of four trp1 gene variants murine pancreatic beta-cells. Diabetologia. 1997 May;40(5):528–532. doi: 10.1007/s001250050711. [DOI] [PubMed] [Google Scholar]
  37. Shieh B. H., Zhu M. Y. Regulation of the TRP Ca2+ channel by INAD in Drosophila photoreceptors. Neuron. 1996 May;16(5):991–998. doi: 10.1016/s0896-6273(00)80122-1. [DOI] [PubMed] [Google Scholar]
  38. Sinkins W. G., Vaca L., Hu Y., Kunze D. L., Schilling W. P. The COOH-terminal domain of Drosophila TRP channels confers thapsigargin sensitivity. J Biol Chem. 1996 Feb 9;271(6):2955–2960. doi: 10.1074/jbc.271.6.2955. [DOI] [PubMed] [Google Scholar]
  39. Takemura H., Putney J. W., Jr Capacitative calcium entry in parotid acinar cells. Biochem J. 1989 Mar 1;258(2):409–412. doi: 10.1042/bj2580409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Thompson J. D., Higgins D. G., Gibson T. J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 1994 Nov 11;22(22):4673–4680. doi: 10.1093/nar/22.22.4673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Tsunoda S., Sierralta J., Sun Y., Bodner R., Suzuki E., Becker A., Socolich M., Zuker C. S. A multivalent PDZ-domain protein assembles signalling complexes in a G-protein-coupled cascade. Nature. 1997 Jul 17;388(6639):243–249. doi: 10.1038/40805. [DOI] [PubMed] [Google Scholar]
  42. Vaca L., Sinkins W. G., Hu Y., Kunze D. L., Schilling W. P. Activation of recombinant trp by thapsigargin in Sf9 insect cells. Am J Physiol. 1994 Nov;267(5 Pt 1):C1501–C1505. doi: 10.1152/ajpcell.1994.267.5.C1501. [DOI] [PubMed] [Google Scholar]
  43. Wes P. D., Chevesich J., Jeromin A., Rosenberg C., Stetten G., Montell C. TRPC1, a human homolog of a Drosophila store-operated channel. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9652–9656. doi: 10.1073/pnas.92.21.9652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Wissenbach U., Schroth G., Philipp S., Flockerzi V. Structure and mRNA expression of a bovine trp homologue related to mammalian trp2 transcripts. FEBS Lett. 1998 Jun 5;429(1):61–66. doi: 10.1016/s0014-5793(98)00561-4. [DOI] [PubMed] [Google Scholar]
  45. Xu X. Z., Li H. S., Guggino W. B., Montell C. Coassembly of TRP and TRPL produces a distinct store-operated conductance. Cell. 1997 Jun 27;89(7):1155–1164. doi: 10.1016/s0092-8674(00)80302-5. [DOI] [PubMed] [Google Scholar]
  46. Zhu X., Jiang M., Birnbaumer L. Receptor-activated Ca2+ influx via human Trp3 stably expressed in human embryonic kidney (HEK)293 cells. Evidence for a non-capacitative Ca2+ entry. J Biol Chem. 1998 Jan 2;273(1):133–142. doi: 10.1074/jbc.273.1.133. [DOI] [PubMed] [Google Scholar]
  47. Zhu X., Jiang M., Peyton M., Boulay G., Hurst R., Stefani E., Birnbaumer L. trp, a novel mammalian gene family essential for agonist-activated capacitative Ca2+ entry. Cell. 1996 May 31;85(5):661–671. doi: 10.1016/s0092-8674(00)81233-7. [DOI] [PubMed] [Google Scholar]
  48. Zitt C., Obukhov A. G., Strübing C., Zobel A., Kalkbrenner F., Lückhoff A., Schultz G. Expression of TRPC3 in Chinese hamster ovary cells results in calcium-activated cation currents not related to store depletion. J Cell Biol. 1997 Sep 22;138(6):1333–1341. doi: 10.1083/jcb.138.6.1333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Zitt C., Zobel A., Obukhov A. G., Harteneck C., Kalkbrenner F., Lückhoff A., Schultz G. Cloning and functional expression of a human Ca2+-permeable cation channel activated by calcium store depletion. Neuron. 1996 Jun;16(6):1189–1196. doi: 10.1016/s0896-6273(00)80145-2. [DOI] [PubMed] [Google Scholar]
  50. Zweifach A., Lewis R. S. Mitogen-regulated Ca2+ current of T lymphocytes is activated by depletion of intracellular Ca2+ stores. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6295–6299. doi: 10.1073/pnas.90.13.6295. [DOI] [PMC free article] [PubMed] [Google Scholar]

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