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Using Nanodiscs to Probe Ca 2+ -Dependent Membrane Interaction of Synaptotagmin-1.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2019; Vol. 1860, pp. 221-236. - Publication Year :
- 2019
-
Abstract
- In this chapter, we introduce a nanodisc-based experimental platform to study Ca <superscript>2+</superscript> -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 Ca <superscript>2+</superscript> -activation process on a millisecond timescale.
- Subjects :
- Calcium metabolism
Cysteine genetics
Fluorescent Dyes chemistry
Lipid Bilayers chemistry
Membrane Fusion
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins isolation & purification
Recombinant Proteins metabolism
Spectrum Analysis instrumentation
Spectrum Analysis methods
Synaptosomal-Associated Protein 25 chemistry
Synaptosomal-Associated Protein 25 isolation & purification
Synaptosomal-Associated Protein 25 metabolism
Synaptotagmin I chemistry
Synaptotagmin I genetics
Synaptotagmin I isolation & purification
Syntaxin 1 chemistry
Syntaxin 1 isolation & purification
Syntaxin 1 metabolism
Lipid Bilayers metabolism
Nanostructures chemistry
Synaptotagmin I metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1860
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 30317508
- Full Text :
- https://doi.org/10.1007/978-1-4939-8760-3_14