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Using Nanodiscs to Probe Ca2+-Dependent Membrane Interaction of Synaptotagmin-1

Ekaterina Stroeva,Shyam S. Krishnakumar

Methods in molecular biology (Clifton, N.J.)（2018）

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Abstract
In this chapter, we introduce a nanodisc-based experimental platform to study Ca2+-triggered membrane interaction of synaptotagmin-1. We describe and discuss in detail how to assemble this soluble mimetic of the docked vesicle-plasma membrane junction, with fluorescently labeled synaptotagmin-1 bound to trans SNAREpins assembled between nanodiscs and present the stopped-flow rapid mixing method used to monitor the conformational dynamics of Ca2+-activation process on a millisecond timescale.
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Calcium activation,Fluorescent labeling,Membrane interaction,Nanodiscs,Rapid mixing stopped-flow,SNARE proteins,Soluble membrane-mimetic,Synaptotagmin-1
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