Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents

This Article Full Text Full Text (PDF, 269K) PPT slides of all figures Alert me when this article is cited Citation Map Services Email this article Similar articles in this journal Similar articles in PubMed Alert me to new content in the JCB Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Earles, C. A. Articles by Chapman, E. R. Search for Related Content PubMed PubMed Citation Articles by Earles, C. A. Articles by Chapman, E. R. Social Bookmarking                            What's this?

© The Rockefeller University Press, 0021-9525/2001/9/1117 $5.00
The Journal of Cell Biology, Volume 154, Number 6, September 17, 2001 1117-1124

The tandem C2 domains of synaptotagmin contain redundant Ca²+ binding sites that cooperate to engage t-SNAREs and trigger exocytosis

Department of Physiology, University of Wisconsin, Madison, WI 53706

Address correspondence to Edwin R. Chapman, Dept. of Physiology, SMI 129, University of Wisconsin, 1300 University Ave., Madison, WI 53706. Tel.: (608) 263-1762. Fax: (608) 265-5512. E-mail: chapman{at}physiology.wisc.edu

Real-time voltammetry measurements from cracked PC12 cells were used to analyze the role of synaptotagmin–SNARE interactions during Ca²+-triggered exocytosis. The isolated C2A domain of synaptotagmin I neither binds SNAREs nor inhibits norepinephrine secretion. In contrast, two C2 domains in tandem (either C2A-C2B or C2A-C2A) bind strongly to SNAREs, displace native synaptotagmin from SNARE complexes, and rapidly inhibit exocytosis. The tandem C2 domains of synaptotagmin cooperate via a novel mechanism in which the disruptive effects of Ca²⁺ ligand mutations in one C2 domain can be partially alleviated by the presence of an adjacent C2 domain. Complete disruption of Ca²⁺-triggered membrane and target membrane SNARE interactions required simultaneous neutralization of Ca²⁺ ligands in both C2 domains of the protein. We conclude that synaptotagmin–SNARE interactions regulate membrane fusion and that cooperation between synaptotagmin's C2 domains is crucial to its function.

Key Words: synaptotagmin; SNARE; membrane fusion; C2 domain; exocytosis

CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Lynch, K. L., Gerona, R. R.L., Kielar, D. M., Martens, S., McMahon, H. T., Martin, T. F.J. (2008). Synaptotagmin-1 Utilizes Membrane Bending and SNARE Binding to Drive Fusion Pore Expansion. Mol. Biol. Cell 19: 5093-5103 [Abstract] [Full Text]  
• Bhalla, A., Chicka, M. C., Chapman, E. R. (2008). Analysis of the Synaptotagmin Family during Reconstituted Membrane Fusion: UNCOVERING A CLASS OF INHIBITORY ISOFORMS. J. Biol. Chem. 283: 21799-21807 [Abstract] [Full Text]  
• Paddock, B. E., Striegel, A. R., Hui, E., Chapman, E. R., Reist, N. E. (2008). Ca2+-Dependent, Phospholipid-Binding Residues of Synaptotagmin Are Critical for Excitation-Secretion Coupling In Vivo. J. Neurosci. 28: 7458-7466 [Abstract] [Full Text]  
• Zhang, Z., Jackson, M. B. (2008). Temperature Dependence of Fusion Kinetics and Fusion Pores in Ca2+-triggered Exocytosis from PC12 Cells. JGP 131: 117-124 [Abstract] [Full Text]  
• Lynch, K. L., Gerona, R.R.L., Larsen, E. C., Marcia, R. F., Mitchell, J. C., Martin, T.F.J. (2007). Synaptotagmin C2A Loop 2 Mediates Ca2+-dependent SNARE Interactions Essential for Ca2+-triggered Vesicle Exocytosis. Mol. Biol. Cell 18: 4957-4968 [Abstract] [Full Text]  
• Gattegno, T., Mittal, A., Valansi, C., Nguyen, K. C.Q., Hall, D. H., Chernomordik, L. V., Podbilewicz, B. (2007). Genetic Control of Fusion Pore Expansion in the Epidermis of Caenorhabditis elegans. Mol. Biol. Cell 18: 1153-1166 [Abstract] [Full Text]  
• Czibener, C., Sherer, N. M., Becker, S. M., Pypaert, M., Hui, E., Chapman, E. R., Mothes, W., Andrews, N. W. (2006). Ca2+ and synaptotagmin VII-dependent delivery of lysosomal membrane to nascent phagosomes. JCB 174: 997-1007 [Abstract] [Full Text]  
• Aganna, E., Burrin, J. M, Hitman, G. A, Turner, M. D (2006). Involvement of calpain and synaptotagmin Ca2+ sensors in hormone secretion from excitable endocrine cells.. J Endocrinol 190: R1-R7 [Abstract] [Full Text]  
• Ahras, M., Otto, G. P., Tooze, S. A. (2006). Synaptotagmin IV is necessary for the maturation of secretory granules in PC12 cells. JCB 173: 241-251 [Abstract] [Full Text]  
• Wang, C.-T., Bai, J., Chang, P. Y., Chapman, E. R., Jackson, M. B. (2006). Synaptotagmin-Ca2+ triggers two sequential steps in regulated exocytosis in rat PC12 cells: fusion pore opening and fusion pore dilation. J. Physiol. 570: 295-307 [Abstract] [Full Text]  
• Rhee, J.-S., Li, L. Y., Shin, O.-H., Rah, J.-C., Rizo, J., Sudhof, T. C., Rosenmund, C. (2005). Augmenting neurotransmitter release by enhancing the apparent Ca2+ affinity of synaptotagmin 1. Proc. Natl. Acad. Sci. USA 102: 18664-18669 [Abstract] [Full Text]  
• Wang, P., Chicka, M. C., Bhalla, A., Richards, D. A., Chapman, E. R. (2005). Synaptotagmin VII Is Targeted to Secretory Organelles in PC12 Cells, Where It Functions as a High-Affinity Calcium Sensor. Mol. Cell. Biol. 25: 8693-8702 [Abstract] [Full Text]  
• Bhalla, A., Tucker, W. C., Chapman, E. R. (2005). Synaptotagmin Isoforms Couple Distinct Ranges of Ca2+, Ba2+, and Sr2+ Concentration to SNARE-mediated Membrane Fusion. Mol. Biol. Cell 16: 4755-4764 [Abstract] [Full Text]  
• DeBlasio, S. L., Luesse, D. L., Hangarter, R. P. (2005). A Plant-Specific Protein Essential for Blue-Light-Induced Chloroplast Movements. Plant Physiol. 139: 101-114 [Abstract] [Full Text]  
• Shen, S. S., Tucker, W. C., Chapman, E. R., Steinhardt, R. A. (2005). Molecular Regulation of Membrane Resealing in 3T3 Fibroblasts. J. Biol. Chem. 280: 1652-1660 [Abstract] [Full Text]  
• Maher, B. J., MacKinnon, R. L. II, Bai, J., Chapman, E. R., Kelly, P. T. (2005). Activation of Postsynaptic Ca2+ Stores Modulates Glutamate Receptor Cycling in Hippocampal Neurons. J. Neurophysiol. 93: 178-188 [Abstract] [Full Text]  
• Llinas, R. R., Sugimori, M., Moran, K. A., Moreira, J. E., Fukuda, M. (2004). Vesicular reuptake inhibition by a synaptotagmin I C2B domain antibody at the squid giant synapse. Proc. Natl. Acad. Sci. USA 101: 17855-17860 [Abstract] [Full Text]  
• Yoshihara, M., Montana, E. S. (2004). The Synaptotagmins: Calcium Sensors for Vesicular Trafficking. Neuroscientist 10: 566-574 [Abstract]  
• Tucker, W. C., Weber, T., Chapman, E. R. (2004). Reconstitution of Ca2+-Regulated Membrane Fusion by Synaptotagmin and SNAREs. Science 304: 435-438 [Abstract] [Full Text]  
• Chieregatti, E., Chicka, M. C., Chapman, E. R., Baldini, G. (2004). SNAP-23 Functions in Docking/Fusion of Granules at Low Ca2+. Mol. Biol. Cell 15: 1918-1930 [Abstract] [Full Text]  
• Wang, P., Wang, C.-T., Bai, J., Jackson, M. B., Chapman, E. R. (2003). Mutations in the Effector Binding Loops in the C2A and C2B Domains of Synaptotagmin I Disrupt Exocytosis in a Nonadditive Manner. J. Biol. Chem. 278: 47030-47037 [Abstract] [Full Text]  
• Edwardson, J. M., Wang, C.-T., Gong, B., Wyttenbach, A., Bai, J., Jackson, M. B., Chapman, E. R., Morton, A. J. (2003). Expression of Mutant Huntingtin Blocks Exocytosis in PC12 Cells by Depletion of Complexin II. J. Biol. Chem. 278: 30849-30853 [Abstract] [Full Text]  
• Duman, J. G., Forte, J. G. (2003). What is the role of SNARE proteins in membrane fusion?. Am. J. Physiol. Cell Physiol. 285: C237-C249 [Abstract] [Full Text]  
• Tucker, W. C., Edwardson, J. M., Bai, J., Kim, H.-J., Martin, T. F.J., Chapman, E. R. (2003). Identification of synaptotagmin effectors via acute inhibition of secretion from cracked PC12 cells. JCB 162: 199-209 [Abstract] [Full Text]  
• Wu, Y., He, Y., Bai, J., Ji, S.-R., Tucker, W. C., Chapman, E. R., Sui, S.-F. (2003). Visualization of synaptotagmin I oligomers assembled onto lipid monolayers. Proc. Natl. Acad. Sci. USA 100: 2082-2087 [Abstract] [Full Text]  
• Fukuda, M., Kanno, E., Ogata, Y., Saegusa, C., Kim, T., Loh, Y. P., Yamamoto, A. (2003). Nerve Growth Factor-dependent Sorting of Synaptotagmin IV Protein to Mature Dense-core Vesicles That Undergo Calcium-dependent Exocytosis in PC12 Cells. J. Biol. Chem. 278: 3220-3226 [Abstract] [Full Text]  
• Fernandez-Chacon, R., Shin, O.-H., Konigstorfer, A., Matos, M. F., Meyer, A. C., Garcia, J., Gerber, S. H., Rizo, J., Sudhof, T. C., Rosenmund, C. (2002). Structure/Function Analysis of Ca2+ Binding to the C2A Domain of Synaptotagmin 1. J. Neurosci. 22: 8438-8446 [Abstract] [Full Text]  
• Fukuda, M. (2002). Vesicle-associated Membrane Protein-2/Synaptobrevin Binding to Synaptotagmin I Promotes O-Glycosylation of Synaptotagmin I. J. Biol. Chem. 277: 30351-30358 [Abstract] [Full Text]  
• Mahal, L. K., Sequeira, S. M., Gureasko, J. M., Sollner, T. H. (2002). Calcium-independent stimulation of membrane fusion and SNAREpin formation by synaptotagmin I. JCB 158: 273-282 [Abstract] [Full Text]  
• Saegusa, C., Fukuda, M., Mikoshiba, K. (2002). Synaptotagmin V Is Targeted to Dense-core Vesicles That Undergo Calcium-dependent Exocytosis in PC12 Cells. J. Biol. Chem. 277: 24499-24505 [Abstract] [Full Text]  
• Honda, A., Yamada, M., Saisu, H., Takahashi, H., Mori, K. J., Abe, T. (2002). Direct, Ca2+-dependent Interaction between Tubulin and Synaptotagmin I. A POSSIBLE MECHANISM FOR ATTACHING SYNAPTIC VESICLES TO MICROTUBULES. J. Biol. Chem. 277: 20234-20242 [Abstract] [Full Text]  
• Bai, J., Wang, P., Chapman, E. R. (2002). C2A activates a cryptic Ca2+-triggered membrane penetration activity within the C2B domain of synaptotagmin I. Proc. Natl. Acad. Sci. USA 10.1073/pnas.032541099v1 [Abstract] [Full Text]  
• Bai, J., Wang, P., Chapman, E. R. (2002). C2A activates a cryptic Ca2+-triggered membrane penetration activity within the C2B domain of synaptotagmin I. Proc. Natl. Acad. Sci. USA 99: 1665-1670 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents