Synaptotagmin oligomers are necessary and can be sufficient to form a Ca2+ -sensitive fusion clamp

Sathish Ramakrishnan, Manindra Bera, Jeff Coleman, Shyam S. Krishnakumar, Frederic Pincet, James E. Rothman
2019 FEBS Letters  
The buttressed-ring hypothesis, supported by recent cryo-electron tomography analysis of docked synaptic-like vesicles in neuroendocrine cells, postulates that prefusion SNAREpins are stabilized and organized by Synaptotagmin (Syt) ring-like oligomers. Here, we use a reconstituted single-vesicle fusion analysis to test the prediction that destabilizing the Syt1 oligomers destabilizes the clamp and results in spontaneous fusion in the absence of Ca 2+ . Vesicles in which Syt oligomerization is
more » ... mpromised by a ring-destabilizing mutation dock and diffuse freely on the bilayer until they fuse spontaneously, similar to vesicles containing only v-SNAREs. In contrast, vesicles containing wild-type Syt are immobile as soon as they attach to the bilayer and remain frozen in place, up to at least 1 h until fusion is triggered by Ca 2+ .
doi:10.1002/1873-3468.13317 fatcat:hpher5yd4jhnxituogstx2kpzu