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. 1996 Jul 9;93(14):7363–7368. doi: 10.1073/pnas.93.14.7363

Isoform-specific interaction of the alpha1A subunits of brain Ca2+ channels with the presynaptic proteins syntaxin and SNAP-25.

J Rettig 1, Z H Sheng 1, D K Kim 1, C D Hodson 1, T P Snutch 1, W A Catterall 1
PMCID: PMC38990  PMID: 8692999

Abstract

Presynaptic Ca2+ channels are crucial elements in neuronal excitation-secretion coupling. In addition to mediating Ca2+ entry to initiate transmitter release, they are thought to interact directly with proteins of the synaptic vesicle docking/fusion machinery. Here we report isoform-specific, stoichiometric interaction of the BI and rbA isoforms of the alpha1A subunit of P/Q-type Ca2+ channels with the presynaptic membrane proteins syntaxin and SNAP-25 in vitro and in rat brain membranes. The BI isoform binds to both proteins, while only interaction with SNAP-25 can be detected in vitro for the rbA isoform. The synaptic protein interaction ("synprint") site involves two adjacent segments of the intracellular loop connecting domains II and III between amino acid residues 722 and 1036 of the BI sequence. This interaction is competitively blocked by the corresponding region of the N-type Ca2+ channel, indicating that these two channels bind to overlapping regions of syntaxin and SNAP-25. Our results provide a molecular basis for a physical link between Ca2+ influx into nerve terminals and subsequent exocytosis of neurotransmitters at synapses that have presynaptic Ca2+ channels containing alpha1A subunits.

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Selected References

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

  1. Bajjalieh S. M., Scheller R. H. The biochemistry of neurotransmitter secretion. J Biol Chem. 1995 Feb 3;270(5):1971–1974. doi: 10.1074/jbc.270.5.1971. [DOI] [PubMed] [Google Scholar]
  2. Bennett M. K., Calakos N., Scheller R. H. Syntaxin: a synaptic protein implicated in docking of synaptic vesicles at presynaptic active zones. Science. 1992 Jul 10;257(5067):255–259. doi: 10.1126/science.1321498. [DOI] [PubMed] [Google Scholar]
  3. Birnbaumer L., Campbell K. P., Catterall W. A., Harpold M. M., Hofmann F., Horne W. A., Mori Y., Schwartz A., Snutch T. P., Tanabe T. The naming of voltage-gated calcium channels. Neuron. 1994 Sep;13(3):505–506. doi: 10.1016/0896-6273(94)90021-3. [DOI] [PubMed] [Google Scholar]
  4. Calakos N., Bennett M. K., Peterson K. E., Scheller R. H. Protein-protein interactions contributing to the specificity of intracellular vesicular trafficking. Science. 1994 Feb 25;263(5150):1146–1149. doi: 10.1126/science.8108733. [DOI] [PubMed] [Google Scholar]
  5. Chapman E. R., An S., Barton N., Jahn R. SNAP-25, a t-SNARE which binds to both syntaxin and synaptobrevin via domains that may form coiled coils. J Biol Chem. 1994 Nov 4;269(44):27427–27432. [PubMed] [Google Scholar]
  6. Dubel S. J., Starr T. V., Hell J., Ahlijanian M. K., Enyeart J. J., Catterall W. A., Snutch T. P. Molecular cloning of the alpha-1 subunit of an omega-conotoxin-sensitive calcium channel. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5058–5062. doi: 10.1073/pnas.89.11.5058. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hayashi T., McMahon H., Yamasaki S., Binz T., Hata Y., Südhof T. C., Niemann H. Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly. EMBO J. 1994 Nov 1;13(21):5051–5061. doi: 10.1002/j.1460-2075.1994.tb06834.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jahn R., Südhof T. C. Synaptic vesicles and exocytosis. Annu Rev Neurosci. 1994;17:219–246. doi: 10.1146/annurev.ne.17.030194.001251. [DOI] [PubMed] [Google Scholar]
  9. Luebke J. I., Dunlap K., Turner T. J. Multiple calcium channel types control glutamatergic synaptic transmission in the hippocampus. Neuron. 1993 Nov;11(5):895–902. doi: 10.1016/0896-6273(93)90119-c. [DOI] [PubMed] [Google Scholar]
  10. Lupas A., Van Dyke M., Stock J. Predicting coiled coils from protein sequences. Science. 1991 May 24;252(5009):1162–1164. doi: 10.1126/science.252.5009.1162. [DOI] [PubMed] [Google Scholar]
  11. Lévêque C., el Far O., Martin-Moutot N., Sato K., Kato R., Takahashi M., Seagar M. J. Purification of the N-type calcium channel associated with syntaxin and synaptotagmin. A complex implicated in synaptic vesicle exocytosis. J Biol Chem. 1994 Mar 4;269(9):6306–6312. [PubMed] [Google Scholar]
  12. Mori Y., Friedrich T., Kim M. S., Mikami A., Nakai J., Ruth P., Bosse E., Hofmann F., Flockerzi V., Furuichi T. Primary structure and functional expression from complementary DNA of a brain calcium channel. Nature. 1991 Apr 4;350(6317):398–402. doi: 10.1038/350398a0. [DOI] [PubMed] [Google Scholar]
  13. Oguro-Okano M., Griesmann G. E., Wieben E. D., Slaymaker S. J., Snutch T. P., Lennon V. A. Molecular diversity of neuronal-type calcium channels identified in small cell lung carcinoma. Mayo Clin Proc. 1992 Dec;67(12):1150–1159. doi: 10.1016/s0025-6196(12)61144-6. [DOI] [PubMed] [Google Scholar]
  14. Oyler G. A., Higgins G. A., Hart R. A., Battenberg E., Billingsley M., Bloom F. E., Wilson M. C. The identification of a novel synaptosomal-associated protein, SNAP-25, differentially expressed by neuronal subpopulations. J Cell Biol. 1989 Dec;109(6 Pt 1):3039–3052. doi: 10.1083/jcb.109.6.3039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sakurai T., Hell J. W., Woppmann A., Miljanich G. P., Catterall W. A. Immunochemical identification and differential phosphorylation of alternatively spliced forms of the alpha 1A subunit of brain calcium channels. J Biol Chem. 1995 Sep 8;270(36):21234–21242. doi: 10.1074/jbc.270.36.21234. [DOI] [PubMed] [Google Scholar]
  16. Sather W. A., Tanabe T., Zhang J. F., Mori Y., Adams M. E., Tsien R. W. Distinctive biophysical and pharmacological properties of class A (BI) calcium channel alpha 1 subunits. Neuron. 1993 Aug;11(2):291–303. doi: 10.1016/0896-6273(93)90185-t. [DOI] [PubMed] [Google Scholar]
  17. Sheng Z. H., Rettig J., Cook T., Catterall W. A. Calcium-dependent interaction of N-type calcium channels with the synaptic core complex. Nature. 1996 Feb 1;379(6564):451–454. doi: 10.1038/379451a0. [DOI] [PubMed] [Google Scholar]
  18. Sheng Z. H., Rettig J., Takahashi M., Catterall W. A. Identification of a syntaxin-binding site on N-type calcium channels. Neuron. 1994 Dec;13(6):1303–1313. doi: 10.1016/0896-6273(94)90417-0. [DOI] [PubMed] [Google Scholar]
  19. Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
  20. Snutch T. P., Leonard J. P., Gilbert M. M., Lester H. A., Davidson N. Rat brain expresses a heterogeneous family of calcium channels. Proc Natl Acad Sci U S A. 1990 May;87(9):3391–3395. doi: 10.1073/pnas.87.9.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Starr T. V., Prystay W., Snutch T. P. Primary structure of a calcium channel that is highly expressed in the rat cerebellum. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5621–5625. doi: 10.1073/pnas.88.13.5621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Stea A., Tomlinson W. J., Soong T. W., Bourinet E., Dubel S. J., Vincent S. R., Snutch T. P. Localization and functional properties of a rat brain alpha 1A calcium channel reflect similarities to neuronal Q- and P-type channels. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10576–10580. doi: 10.1073/pnas.91.22.10576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Takahashi T., Momiyama A. Different types of calcium channels mediate central synaptic transmission. Nature. 1993 Nov 11;366(6451):156–158. doi: 10.1038/366156a0. [DOI] [PubMed] [Google Scholar]
  24. Turner T. J., Adams M. E., Dunlap K. Multiple Ca2+ channel types coexist to regulate synaptosomal neurotransmitter release. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9518–9522. doi: 10.1073/pnas.90.20.9518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Westenbroek R. E., Sakurai T., Elliott E. M., Hell J. W., Starr T. V., Snutch T. P., Catterall W. A. Immunochemical identification and subcellular distribution of the alpha 1A subunits of brain calcium channels. J Neurosci. 1995 Oct;15(10):6403–6418. doi: 10.1523/JNEUROSCI.15-10-06403.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wheeler D. B., Randall A., Tsien R. W. Roles of N-type and Q-type Ca2+ channels in supporting hippocampal synaptic transmission. Science. 1994 Apr 1;264(5155):107–111. doi: 10.1126/science.7832825. [DOI] [PubMed] [Google Scholar]
  27. Williams M. E., Brust P. F., Feldman D. H., Patthi S., Simerson S., Maroufi A., McCue A. F., Veliçelebi G., Ellis S. B., Harpold M. M. Structure and functional expression of an omega-conotoxin-sensitive human N-type calcium channel. Science. 1992 Jul 17;257(5068):389–395. doi: 10.1126/science.1321501. [DOI] [PubMed] [Google Scholar]
  28. Yoshida A., Oho C., Omori A., Kuwahara R., Ito T., Takahashi M. HPC-1 is associated with synaptotagmin and omega-conotoxin receptor. J Biol Chem. 1992 Dec 15;267(35):24925–24928. [PubMed] [Google Scholar]

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