Single-Molecule Optical Tweezers Study of Regulated SNARE Assembly
| Autor: | Lu Ma, Yongli Zhang, Junyi Jiao |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Protein Folding
Optical Tweezers Membrane Fusion Article Intracellular membrane 03 medical and health sciences 0302 clinical medicine Molecule Biotinylation Protein Structure Quaternary Cellular compartment 030304 developmental biology 0303 health sciences Chemistry Lipid bilayer fusion Energy landscape Microfluidic Analytical Techniques Recombinant Proteins Single Molecule Imaging Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins Folding (chemistry) Kinetics Cross-Linking Reagents Optical tweezers Biophysics Spatiotemporal resolution SNARE Proteins 030217 neurology & neurosurgery Protein Binding |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493987597 |
| Popis: | Intracellular membrane fusion mediates material and information exchange among different cells or cellular compartments with high accuracy and spatiotemporal resolution. Fusion is driven by ordered folding and assembly of soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein (SNAP) receptors (SNAREs) and regulated by many other proteins. Understanding regulated SNARE assembly is key to dissecting mechanisms and physiologies of various fusion processes and their associated diseases. Yet, it remains challenging to study regulated SNARE assembly using traditional ensemble-based experimental approaches. Here, we describe our new method to measure the energy and kinetics of neuronal SNARE assembly in the presence of α-SNAP, using a single-molecule manipulation approach based on high-resolution optical tweezers. Detailed experimental protocols and methods of data analysis are shown. This approach can be widely applied to elucidate the effects of regulatory proteins on SNARE assembly and membrane fusion. |
| Databáze: | OpenAIRE |
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