Dynamin Superfamily GTPases: Methods and Protocols

✍ Scribed by Rajesh Ramachandran

• Publisher: Springer US;Humana
• Year: 2020
• Tongue: English
• Leaves: 219
• Series: Methods in Molecular Biology 2159
• Edition: 1st ed.
• Category: Library

✦ Synopsis

This volume provides methods and protocols on dynamin superfamily GTPases. Chapters are divided into three parts; detailing expression, purification, initial biochemical steps, undescribed analytical techniques, and biophysical and cellular characterization of DSPs in membrane remodeling, fission and fusion. 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, Dynamin Superfamily GTPases: Methods and Protocols to ensure successful results in the further study of this vital field.

✦ Table of Contents

Front Matter ....Pages i-x
Front Matter ....Pages 1-1
Isolation and Analysis of Mitochondrial Fission Enzyme DNM1 from Saccharomyces cerevisiae (Nolan W. Kennedy, Lora K. Picton, R. Blake Hill)....Pages 3-15
Purification of the Dynamin-Related Protein Vps1 Using Mammalian and Bacterial Expression Systems (Natalia V. Varlakhanova, Marijn G. J. Ford)....Pages 17-27
Front Matter ....Pages 29-29
A Single Common Protocol for the Expression and Purification of Soluble Mammalian DSPs from Escherichia coli (Natalia Stepanyants, Patrick J. Macdonald, Pooja Madan Mohan, Rajesh Ramachandran)....Pages 31-40
Affinity Purification and Functional Characterization of Dynamin-Related Protein 1 (Ryan W. Clinton, Brianna L. Bauer, Jason A. Mears)....Pages 41-53
Purification and Characterization of MxB (Frances Joan D. Alvarez, Peijun Zhang)....Pages 55-65
Purification of FarnesylatedhGBP1 and Characterization of Its Polymerization and Membrane Binding (Linda Sistemich, Christian Herrmann)....Pages 67-81
Front Matter ....Pages 83-83
Microscale Thermophoresis (MST) as a Tool for Measuring Dynamin Superfamily Protein (DSP)–Lipid Interactions (Nikhil Bharambe, Rajesh Ramachandran)....Pages 85-92
Nucleotide-Dependent Dimerization and Conformational Switching of Atlastin (John P. O’Donnell, Carolyn M. Kelly, Holger Sondermann)....Pages 93-113
Analysis of Mitochondrial Membrane Fusion GTPase OPA1 Expressed by the Silkworm Expression System (Tadato Ban, Naotada Ishihara)....Pages 115-127
Cell-Free Analysis of Mitochondrial Fusion by Fluorescence Microscopy (Nyssa Becker Samanas, Suzanne Hoppins)....Pages 129-140
Electrophysiological Methods for Detection of Membrane Leakage and Hemifission by Dynamin 1 (Pavel V. Bashkirov, Ksenia V. Chekashkina, Anna V. Shnyrova, Vadim A. Frolov)....Pages 141-162
Integrating Optical and Electrochemical Approaches to Assess the Actions of Dynamin at the Fusion Pore (Katherine A. Smith, Emily R. Prantzalos, Arun Anantharam)....Pages 163-178
Cellular Assays for Measuring Dynamin Activity in Muscle Cells (Jessica Laiman, Ya-Wen Liu)....Pages 179-187
Measuring Drp1 Activity in Mitochondrial Fission In Vivo (Di Hu, Xin Qi)....Pages 189-195
Quantifying Drp1-Mediated Mitochondrial Fission by Immunostaining in Fixed Cells (Di Hu, Xin Qi)....Pages 197-204
Imaging Dynamin-Related Protein 1 (Drp1)-Mediated Mitochondrial Fission in Living Cells (Felipe Montecinos-Franjola, Rajesh Ramachandran)....Pages 205-217
Correction to: Imaging Dynamin-Related Protein 1 (Drp1)-Mediated Mitochondrial Fission in Living Cells (Felipe Montecinos-Franjola, Rajesh Ramachandran)....Pages C1-C1
Back Matter ....Pages 219-221

✦ Subjects

Chemistry; Chemistry/Food Science, general