| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Outline of Final Research Achievements |
SNAREs (syntaxin/SNAP-25/synaptobrevin) and synaptotagmin play central roles during Ca2+-dependent neurotransmitter release. In this study, we have studied the effects of synaptotagmin binding on SNARE-mediated fusion and its complex formation. Synaptotagmin stoichiometrically bound to syntaxin in the liposomes containing t-SNAREs (syntaxin, SNAP-25) and dissociated from it by Ca2+. The fusion between synaptobrevin-liposomes and t-SNARE liposomes was inhibited in the presence of synaptotagmin cytoplasmic fragment. This inhibition was reversed by Ca2+. FRET analysis of trans-SNARE complex formation showed that the assembly of SNARE motifs appears to proceed to +6th layers when synaptotagmin binds syntaxin. These results support the idea where synaptotagmin suppresses SNARE-mediated membrane fusion through its syntaxin binding until Ca2+ influx without inhibiting the full assembly of the trans-SNARE complex.
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