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Published online 15 July 2002. doi:10.1083/jcb.200203135
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© The Rockefeller University Press, 0021-9525/2002/7/273 $5.00
The Journal of Cell Biology, Volume 158, Number 2, July 22, 2002 273-282

Article

 Calcium-independent stimulation of membrane fusion and SNAREpin formation by synaptotagmin I

Lara K. Mahal, Sonia M. Sequeira, Jodi M. Gureasko and Thomas H. Söllner

Cellular Biochemistry and Biophysics Program, Memorial Sloan Kettering Cancer Center, New York, NY 10021

Address correspondence to Thomas H. Söllner, Cellular Biochemistry and Biophysics Program, Memorial Sloan Kettering Cancer Center, 1275 York Ave., Rm. 517D, New York, NY 10021. Tel.: (212) 639-5172. Fax: (212) 717-3604. E-mail: t-sollner{at}ski.mskcc.org

Ñeurotransmitter release requires the direct coupling of the calcium sensor with the machinery for membrane fusion. SNARE proteins comprise the minimal fusion machinery, and synaptotagmin I, a synaptic vesicle protein, is the primary candidate for the main neuronal calcium sensor. To test the effect of synaptotagmin I on membrane fusion, we incorporated it into a SNARE-mediated liposome fusion assay. Synaptotagmin I dramatically stimulated membrane fusion by facilitating SNAREpin zippering. This stimulatory effect was topologically restricted to v-SNARE vesicles (containing VAMP 2) and only occurred in trans to t-SNARE vesicles (containing syntaxin 1A and SNAP-25). Interestingly, calcium did not affect the overall fusion reaction. These results indicate that synaptotagmin I can directly accelerate SNARE-mediated membrane fusion and raise the possibility that additional components might be required to ensure tight calcium coupling.

Key Words: SNARE; fusion; synaptotagmin; calcium; exocytosis


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