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- W2912158391 abstract "Neurotransmitter and hormone release involve calcium-triggered fusion of a cargo-loaded vesicle with the plasma membrane. The initial connection between the fusing membranes, called the fusion pore, can evolve in various ways, including rapid dilation to allow full cargo release, slow expansion, repeated opening-closing, and resealing. Pore dynamics determine kinetics of cargo release and mode of vesicle recycling, but how these processes are controlled are poorly understood. Most previous reconstitutions could not monitor single pores, limiting mechanistic insight they could provide into pore regulation. We recently developed a nanodisc-based fusion assay that allow reconstitution and monitoring of single pores with unprecedented detail. The method uses cells expressing “flipped” neuronal SNAREs on their surfaces as fusion partners with nanodiscs reconstituted with the complementary SNAREs. The fusion pore that forms between cell and nanodisc membranes generates a connection between the cell interior and exterior that can be probed by monitoring pore conductance under voltage-clamp. Previously, using this assay we had characterized fusion pores induced by SNAREs alone. Here we studied the role of Synaptotagmin-1 C2AB domains in pore regulation. We found C2AB domains promote pore dilation in a calcium, SNARE, and PI(4,5)P2 dependent manner. Both C2 domains and calcium-induced membrane-insertion of hydrophobic loops at the tips of the calcium-binding loops are important for pore dilation." @default.
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- W2912158391 date "2019-02-01" @default.
- W2912158391 modified "2023-09-30" @default.
- W2912158391 title "Fusion Pore Dilation by Synaptotagmin-1" @default.
- W2912158391 doi "https://doi.org/10.1016/j.bpj.2018.11.2834" @default.
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