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Functional Reconstitution of Intracellular Vesicle Fusion Using Purified SNAREs and Sec1/Munc18 (SM) Proteins

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SNAREs

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

Abstract

The fusion of intracellular vesicles with target membranes is mediated by two classes of conserved molecules—soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAP receptors or SNAREs) and Sec1/Munc18 (SM) proteins. A conserved function of SM proteins is to recognize their cognate trans-SNARE complexes and accelerate fusion kinetics. Here, we describe a physiologically relevant reconstitution system in which macromolecular crowding agents are included to recapitulate the crowded intracellular environment. Through this system, we elucidate the molecular mechanisms by which SNAREs and SM proteins drive vesicle fusion.

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Acknowledgments

This work was supported by National Institutes of Health grants GM102217 and DK095367 (JS).

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Authors and Affiliations

  1. Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing, China

    Haijia Yu

  2. Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, CO, USA

    Haijia Yu, Lauren Crisman, Michael H. B. Stowell & Jingshi Shen

Authors
  1. Haijia Yu
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  2. Lauren Crisman
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  3. Michael H. B. Stowell
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  4. Jingshi Shen
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Corresponding authors

Correspondence to Haijia Yu or Jingshi Shen .

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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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Yu, H., Crisman, L., Stowell, M.H.B., Shen, J. (2019). Functional Reconstitution of Intracellular Vesicle Fusion Using Purified SNAREs and Sec1/Munc18 (SM) Proteins. 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_15

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  • DOI: https://doi.org/10.1007/978-1-4939-8760-3_15

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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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