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Single-Molecule Optical Tweezers Study of Regulated SNARE Assembly

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SNAREs

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1860))

Abstract

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.

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Acknowledgments

This work was supported by NIH grants RO1GM093341 and RO1GM120193 to Y.Z. We also thank the support from the National Natural Science Foundation of China (31770812 to M.L.) and the Chinese Academy of Science Chengkun Wang talent program and Jiaxi Lu international team.

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Authors and Affiliations

  1. Beijing National Laboratory for Condensed Matter Physics and CAS Key Laboratory of Soft Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing, China

    Lu Ma

  2. Department of Cell Biology, Yale University School of Medicine, New Haven, CT, USA

    Junyi Jiao & Yongli Zhang

  3. Integrated Graduate Program in Physical and Engineering Biology, New Haven, CT, USA

    Junyi Jiao

Authors
  1. Lu Ma
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  2. Junyi Jiao
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  3. Yongli Zhang
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Corresponding author

Correspondence to Yongli Zhang .

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Editors and Affiliations

  1. Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, IL, USA

    Rutilio Fratti

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Ma, L., Jiao, J., Zhang, Y. (2019). Single-Molecule Optical Tweezers Study of Regulated SNARE Assembly. In: Fratti, R. (eds) SNAREs. Methods in Molecular Biology, vol 1860. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8760-3_6

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  • DOI: https://doi.org/10.1007/978-1-4939-8760-3_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8759-7

  • Online ISBN: 978-1-4939-8760-3

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