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. 1993 Jul 1;90(13):6228–6232. doi: 10.1073/pnas.90.13.6228

Cloning, chromosomal localization, and functional expression of the alpha 1 subunit of the L-type voltage-dependent calcium channel from normal human heart.

D Schultz 1, G Mikala 1, A Yatani 1, D B Engle 1, D E Iles 1, B Segers 1, R J Sinke 1, D O Weghuis 1, U Klöckner 1, M Wakamori 1, et al.
PMCID: PMC46901  PMID: 8392192

Abstract

A unique structural variant of the cardiac L-type voltage-dependent calcium channel alpha 1 subunit cDNA was isolated from libraries derived from normal human heart mRNA. The deduced amino acid sequence shows significant homology to other calcium channel alpha 1 subunits. However, differences from the rabbit heart alpha 1 include a shortened N-terminus, a unique C-terminal insertion, and both forms of an alternatively spliced motif IV S3 region. The shortened N-terminus provides optimal access to consensus sequences thought to facilitate translation. Northern blot analysis revealed a single hybridizing mRNA species of 9.4 kb. The gene for the human heart alpha 1 subunit was localized specifically to the distal region of chromosome 12p13. The cloned alpha 1 subunit was expressed in Xenopus oocytes and single-channel analyses revealed native-like pharmacology and channel properties.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Biel M., Ruth P., Bosse E., Hullin R., Stühmer W., Flockerzi V., Hofmann F. Primary structure and functional expression of a high voltage activated calcium channel from rabbit lung. FEBS Lett. 1990 Sep 3;269(2):409–412. doi: 10.1016/0014-5793(90)81205-3. [DOI] [PubMed] [Google Scholar]
  2. Catterall W. A. Functional subunit structure of voltage-gated calcium channels. Science. 1991 Sep 27;253(5027):1499–1500. doi: 10.1126/science.1654596. [DOI] [PubMed] [Google Scholar]
  3. Cavalié A., Ochi R., Pelzer D., Trautwein W. Elementary currents through Ca2+ channels in guinea pig myocytes. Pflugers Arch. 1983 Sep;398(4):284–297. doi: 10.1007/BF00657238. [DOI] [PubMed] [Google Scholar]
  4. Chin H. M., Kozak C. A., Kim H. L., Mock B., McBride O. W. A brain L-type calcium channel alpha 1 subunit gene (CCHL1A2) maps to mouse chromosome 14 and human chromosome 3. Genomics. 1991 Dec;11(4):914–919. doi: 10.1016/0888-7543(91)90014-6. [DOI] [PubMed] [Google Scholar]
  5. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  6. Ellis S. B., Williams M. E., Ways N. R., Brenner R., Sharp A. H., Leung A. T., Campbell K. P., McKenna E., Koch W. J., Hui A. Sequence and expression of mRNAs encoding the alpha 1 and alpha 2 subunits of a DHP-sensitive calcium channel. Science. 1988 Sep 23;241(4873):1661–1664. doi: 10.1126/science.2458626. [DOI] [PubMed] [Google Scholar]
  7. Koch W. J., Ellinor P. T., Schwartz A. cDNA cloning of a dihydropyridine-sensitive calcium channel from rat aorta. Evidence for the existence of alternatively spliced forms. J Biol Chem. 1990 Oct 15;265(29):17786–17791. [PubMed] [Google Scholar]
  8. Kozak M. Structural features in eukaryotic mRNAs that modulate the initiation of translation. J Biol Chem. 1991 Oct 25;266(30):19867–19870. [PubMed] [Google Scholar]
  9. Mikami A., Imoto K., Tanabe T., Niidome T., Mori Y., Takeshima H., Narumiya S., Numa S. Primary structure and functional expression of the cardiac dihydropyridine-sensitive calcium channel. Nature. 1989 Jul 20;340(6230):230–233. doi: 10.1038/340230a0. [DOI] [PubMed] [Google Scholar]
  10. Miller S. A., Dykes D. D., Polesky H. F. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988 Feb 11;16(3):1215–1215. doi: 10.1093/nar/16.3.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Perez-Reyes E., Castellano A., Kim H. S., Bertrand P., Baggstrom E., Lacerda A. E., Wei X. Y., Birnbaumer L. Cloning and expression of a cardiac/brain beta subunit of the L-type calcium channel. J Biol Chem. 1992 Jan 25;267(3):1792–1797. [PubMed] [Google Scholar]
  12. Perez-Reyes E., Wei X. Y., Castellano A., Birnbaumer L. Molecular diversity of L-type calcium channels. Evidence for alternative splicing of the transcripts of three non-allelic genes. J Biol Chem. 1990 Nov 25;265(33):20430–20436. [PubMed] [Google Scholar]
  13. Powers P. A., Gregg R. G., Lalley P. A., Liao M., Hogan K. Assignment of the human gene for the alpha 1 subunit of the cardiac DHP-sensitive Ca2+ channel (CCHL1A1) to chromosome 12p12-pter. Genomics. 1991 Jul;10(3):835–839. doi: 10.1016/0888-7543(91)90471-p. [DOI] [PubMed] [Google Scholar]
  14. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sinke R. J., Suijkerbuijk R. F., Herbergs J., Janssen H., Cassiman J. J., Geurts van Kessel A. Generation of a panel of somatic cell hybrids containing fragments of human chromosome 12P by X-ray irradiation and cell fusion. Genomics. 1992 Feb;12(2):206–213. doi: 10.1016/0888-7543(92)90367-2. [DOI] [PubMed] [Google Scholar]
  16. Slish D. F., Engle D. B., Varadi G., Lotan I., Singer D., Dascal N., Schwartz A. Evidence for the existence of a cardiac specific isoform of the alpha 1 subunit of the voltage dependent calcium channel. FEBS Lett. 1989 Jul 3;250(2):509–514. doi: 10.1016/0014-5793(89)80786-0. [DOI] [PubMed] [Google Scholar]
  17. Snutch T. P., Tomlinson W. J., Leonard J. P., Gilbert M. M. Distinct calcium channels are generated by alternative splicing and are differentially expressed in the mammalian CNS. Neuron. 1991 Jul;7(1):45–57. doi: 10.1016/0896-6273(91)90073-9. [DOI] [PubMed] [Google Scholar]
  18. Soldatov N. M. Molecular diversity of L-type Ca2+ channel transcripts in human fibroblasts. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4628–4632. doi: 10.1073/pnas.89.10.4628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Suijkerbuijk R. F., Matthopoulos D., Kearney L., Monard S., Dhut S., Cotter F. E., Herbergs J., van Kessel A. G., Young B. D. Fluorescent in situ identification of human marker chromosomes using flow sorting and Alu element-mediated PCR. Genomics. 1992 Jun;13(2):355–362. doi: 10.1016/0888-7543(92)90253-o. [DOI] [PubMed] [Google Scholar]
  20. Tanabe T., Takeshima H., Mikami A., Flockerzi V., Takahashi H., Kangawa K., Kojima M., Matsuo H., Hirose T., Numa S. Primary structure of the receptor for calcium channel blockers from skeletal muscle. Nature. 1987 Jul 23;328(6128):313–318. doi: 10.1038/328313a0. [DOI] [PubMed] [Google Scholar]
  21. Williams J. S., Grupp I. L., Grupp G., Vaghy P. L., Dumont L., Schwartz A., Yatani A., Hamilton S., Brown A. M. Profile of the oppositely acting enantiomers of the dihydropyridine 202-791 in cardiac preparations: receptor binding, electrophysiological, and pharmacological studies. Biochem Biophys Res Commun. 1985 Aug 30;131(1):13–21. doi: 10.1016/0006-291x(85)91763-2. [DOI] [PubMed] [Google Scholar]
  22. Yatani A., Codina J., Imoto Y., Reeves J. P., Birnbaumer L., Brown A. M. A G protein directly regulates mammalian cardiac calcium channels. Science. 1987 Nov 27;238(4831):1288–1292. doi: 10.1126/science.2446390. [DOI] [PubMed] [Google Scholar]

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