SNAREs: Methods and Protocols

Front Matter ....Pages i-xiv
Front Matter ....Pages 1-1
 Molecular Dynamics Simulations of the SNARE Complex  (Maria Bykhovskaia)....Pages 3-13
 Mesoscale Computational Modeling of Protein-Membrane Interactions Based on Continuum Mean-Field Theory  (George Khelashvili)....Pages 15-31
 EPR Lineshape Analysis to Investigate the SNARE Folding Intermediates  (Ryan Khounlo, Brenden J. D. Hawk, Yeon-Kyun Shin)....Pages 33-51
 Dynamic Light Scattering Analysis to Dissect Intermediates of SNARE-Mediated Membrane Fusion  (Byoungjae Kong, Yoosoo Yang, Dae-Hyuk Kweon)....Pages 53-69
 SNAREpin Assembly: Kinetic and Thermodynamic Approaches  (Feng Li, Frederic Pincet)....Pages 71-93
 Single-Molecule Optical Tweezers Study of Regulated SNARE Assembly  (Lu Ma, Junyi Jiao, Yongli Zhang)....Pages 95-114
 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
 Using Force Spectroscopy to Probe Coiled-Coil Assembly and Membrane Fusion  (Hannes Witt, Andreas Janshoff)....Pages 145-159
Front Matter ....Pages 161-161
 SNAP-25 S-Guanylation and SNARE Complex Formation  (Yusuke Kishimoto, Takaaki Akaike, Hideshi Ihara)....Pages 163-173
 Analysis of the Role of Sec3 in SNARE Assembly and Membrane Fusion  (Kunrong Mei, Peng Yue, Wei Guo)....Pages 175-189
 Use of Microscale Thermophoresis (MST) to Measure Binding Affinities of Components of the Fusion Machinery  (Robert P. Sparks, Rutilio Fratti)....Pages 191-198
 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
 Determination of Sec18-Lipid Interactions by Liposome-Binding Assay  (Matthew L. Starr, Rutilio Fratti)....Pages 211-220
 Using Nanodiscs to Probe Ca2+-Dependent Membrane Interaction of Synaptotagmin-1  (Ekaterina Stroeva, Shyam S. Krishnakumar)....Pages 221-236
 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
Front Matter ....Pages 251-251
 Assay of Lipid Mixing and Fusion Pore Formation in the Fusion of Yeast Vacuoles  (Massimo D’Agostino, Andreas Mayer)....Pages 253-262
 A Nanodisc-Cell Fusion Assay with Single-Pore Sensitivity and Sub-millisecond Time Resolution  (Natasha R. Dudzinski, Zhenyong Wu, Erdem Karatekin)....Pages 263-275
 An In Vitro Assay of Trans-SNARE Complex Formation During Yeast Vacuole Fusion Using Epitope Tag-Free SNAREs  (Youngsoo Jun)....Pages 277-288
 A Cell-Free Content Mixing Assay for SNARE-Mediated Multivesicular Body-Vacuole Membrane Fusion  (Mahmoud Abdul Karim, Dieter Ronny Samyn, Christopher Leonard Brett)....Pages 289-301
 Reconstituted Proteoliposome Fusion Mediated by Yeast SNARE-Family Proteins  (Joji Mima)....Pages 303-322
 Real-Time Fluorescence Detection of Calcium Efflux During Vacuolar Membrane Fusion  (Gregory E. Miner, Rutilio Fratti)....Pages 323-331
Front Matter ....Pages 333-333
 Single-Molecule Fluorescence Measurement of SNARE-Mediated Vesicle Fusion  (Yachong Hu, Zhiqi Tian, Jiajie Diao)....Pages 335-344
 Quantifying Intramolecular Protein Conformational Dynamics Under Lipid Interaction Using smFRET and FCCS  (Pei Li, Yawei Dai, Markus Seeger, Yan-Wen Tan)....Pages 345-359
 Visualization of SNARE-Mediated Organelle Membrane Hemifusion by Electron Microscopy  (Sevan Mattie, Tom Kazmirchuk, Jeannie Mui, Hojatollah Vali, Christopher Leonard Brett)....Pages 361-377
 Studies of the Secretory Machinery Dynamics by Total Internal Reflection Fluorescence Microscopy in Bovine Adrenal Chromaffin Cells  (José Villanueva, Yolanda Gimenez-Molina, Luis M. Gutiérrez)....Pages 379-389
 Imaging SNAP-29 in Drosophila  (Hao Xu, Bryan Stewart)....Pages 391-401
Back Matter ....Pages 403-407