PART I: Yeast DSPs – Isolation and Biochemical Characterization
1. Isolation and Analysis of Mitochondrial Fission Enzyme DNM1 from Saccharomyces cerevisiae
Nolan W. Kennedy, Lora K. Picton, and R. Blake Hill
2. Purification of the Dynamin-related Protein Vps1 using Mammalian and Bacterial Expression Systems
Natalia V. Varlakhanovaand Marijn G. J. Ford
PART II: Mammalian DSPs – Isolation and Biochemical Characterization
3. A Single Common Protocol for the Expression and Purification of Soluble Mammalian DSPs from Escherichia coli
Natalia Stepanyants, Patrick J. Macdonald, Pooja Madan Mohan, and Rajesh Ramachandran
4. Affinity Purification and Functional Characterization of Dynamin-related Protein 1
Ryan W. Clinton, Brianna L. Bauer, and Jason A. Mears
5. Purification and Characterization of MxB
Frances Joan D. Alvarez and Peijun Zhang
6. Purification of Farnesylated hGBP1 and Characterization of Its Polymerization and Membrane Binding
Linda Sistemich and Christian Herrmann
PART III: Biophysical and Cellular Characterization of DSPs
7. Microscale Thermophoresis (MST) as a Tool for Measuring Dynamin Superfamily Protein (DSP)-lipid Interactions
Nikhil Bharambe and Rajesh Ramachandran
8. Nucleotide-dependent Dimerization and Conformational Switching of Atlastin
John P. O’Donnell, Carolyn M. Kelly, and Holger Sondermann
9. Analysis of Mitochondrial Membrane Fusion GTPase OPA1 Expressed by the Silkworm Expression System
Tadato Ban and Naotada Ishihara
10. Cell-free Analysis of Mitochondrial Fusion by Fluorescence Microscopy
Nyssa Becker Samanas and Suzanne Hoppins
11. Electrophysiological Methods For Detection Of Membrane Leakage and Hemifission by Dynamin 1
Pavel V. Bashkirov, Ksenia V. Chekashkina, Anna V. Shnyrova, and Vadim A. Frolov
12. Integrating Optical and Electrochemical Approaches to Assess the Actions of Dynamin at the Fusion Pore
Katherine A. Smith, Emily R. Prantzalos, and Arun Anantharam
13. Cellular Assays for Measuring Dynamin Activity in Muscle Cells
Jessica Laimanand Ya-Wen Liu
14. Measuring Drp1 Activity in Mitochondrial Fission in vivo
Di Hu and Xin Qi
15. Quantifying Drp1-mediated Mitochondrial Fission by Immunostaining in Fixed Cells
Di Hu and Xin Qi
16. Imaging Dynamin-related Protein 1 (Drp1)-mediated Mitochondrial Fission in Living Cells
Felipe Montecinos-Franjola and Rajesh Ramachandran
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.