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SNAREs

Methods and Protocols

  • Book
  • © 2019

Overview

Editors:
  1. Rutilio Fratti

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

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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

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

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About this book

This book details multiple ways that soluble N-ethylmaleimide-sensitive factor attachment protein receptors( SNAREs) and their function are examined in the laboratory. The methods described in each chapter are described in detail so that novice as well as experienced researchers can explore the mechanisms of SNARE-mediated membrane fusion. Chapters guide readers through an overview of the basic properties of SNAREs, distribution and interaction with regulators of membrane fusion, activation of SNAREs in the priming stage by NSF/Sec18 and a-SNAP/Sec17, examining the structures and interactions of SNAREs, measuring the interactions of SNAREs, interactions of SNAREs, and post-translational modifications of SNAREs and how they affect function. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, SNAREs: Methods and Protocols aims to be a valuable tool for all investigators interested in the field.

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Keywords

Table of contents (26 protocols)

  1. Front Matter

    Pages i-xiv
    Download chapter PDF

  2. Biophysics and Computational Analysis

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. Molecular Dynamics Simulations of the SNARE Complex

      • Maria Bykhovskaia
      Pages 3-13

    3. Mesoscale Computational Modeling of Protein-Membrane Interactions Based on Continuum Mean-Field Theory

      • George Khelashvili
      Pages 15-31

    4. EPR Lineshape Analysis to Investigate the SNARE Folding Intermediates

      • Ryan Khounlo, Brenden J. D. Hawk, Yeon-Kyun Shin
      Pages 33-51

    5. Dynamic Light Scattering Analysis to Dissect Intermediates of SNARE-Mediated Membrane Fusion

      • Byoungjae Kong, Yoosoo Yang, Dae-Hyuk Kweon
      Pages 53-69

    6. SNAREpin Assembly: Kinetic and Thermodynamic Approaches

      • Feng Li, Frederic Pincet
      Pages 71-93

    7. Single-Molecule Optical Tweezers Study of Regulated SNARE Assembly

      • Lu Ma, Junyi Jiao, Yongli Zhang
      Pages 95-114

    8. Studying Munc18:Syntaxin Interactions Using Small-Angle Scattering

      • Andrew E. Whitten, Russell J. Jarrott, Shu-Hong Hu, Anthony P. Duff, Gordon J. King, Jennifer L. Martin et al.
      Pages 115-144

    9. Using Force Spectroscopy to Probe Coiled-Coil Assembly and Membrane Fusion

      • Hannes Witt, Andreas Janshoff
      Pages 145-159

  3. Biochemistry

    1. Front Matter

      Pages 161-161
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    2. SNAP-25 S-Guanylation and SNARE Complex Formation

      • Yusuke Kishimoto, Takaaki Akaike, Hideshi Ihara
      Pages 163-173

    3. Analysis of the Role of Sec3 in SNARE Assembly and Membrane Fusion

      • Kunrong Mei, Peng Yue, Wei Guo
      Pages 175-189

    4. Use of Microscale Thermophoresis (MST) to Measure Binding Affinities of Components of the Fusion Machinery

      • Robert P. Sparks, Rutilio Fratti
      Pages 191-198

    5. Use of Surface Plasmon Resonance (SPR) to Determine Binding Affinities and Kinetic Parameters Between Components Important in Fusion Machinery

      • Robert P. Sparks, Jermaine L. Jenkins, Rutilio Fratti
      Pages 199-210

    6. Determination of Sec18-Lipid Interactions by Liposome-Binding Assay

      • Matthew L. Starr, Rutilio Fratti
      Pages 211-220

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

      • Ekaterina Stroeva, Shyam S. Krishnakumar
      Pages 221-236

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

      • Haijia Yu, Lauren Crisman, Michael H. B. Stowell, Jingshi Shen
      Pages 237-249

  4. Functional Assays

    1. Front Matter

      Pages 251-251
      Download chapter PDF

    2. Assay of Lipid Mixing and Fusion Pore Formation in the Fusion of Yeast Vacuoles

      • Massimo D’Agostino, Andreas Mayer
      Pages 253-262
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Editors and Affiliations

  • Department of Biochemistry, University of Illinois Urbana-Champaign, Urbana, USA

    Rutilio Fratti

Bibliographic Information

  • Book Title: SNAREs

  • Book Subtitle: Methods and Protocols

  • Editors: Rutilio Fratti

  • Series Title: Methods in Molecular Biology

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

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2019

  • Hardcover ISBN: 978-1-4939-8759-7Published: 14 October 2018

  • Softcover ISBN: 978-1-4939-9385-7Published: 10 December 2019

  • eBook ISBN: 978-1-4939-8760-3Published: 13 October 2018

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIV, 407

  • Number of Illustrations: 8 b/w illustrations, 66 illustrations in colour

  • Topics: Biochemistry, general

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