The ESCRT Complexes: Methods and Protocols

✍ Scribed by Emmanuel Culetto, Renaud Legouis

• Publisher: Springer New York; Humana
• Year: 2019
• Tongue: English
• Leaves: 327
• Series: Methods in Molecular Biology 1998
• Edition: 1st ed.
• Category: Library

✦ Synopsis

This detailed collection gathers both established and recent technical procedures to study the Endosomal Sorting Complex Required for Transport (ESCRT) complexes in a wide range of biological systems: Archaea, A. thaliana, U. maydis, S. cerevisiae, S. pombe, C. elegans, D. melanogaster, and mammalian cells. Opening with a section on imaging techniques, the book continues with chapters covering biochemical approaches presenting strategies for production and characterization of recombinant ESCRT proteins, or of specific ESCRT protein domains, as well as genetic and proteomic experimental approaches. Written for the highly successful Methods in Molecular Biology series, chapters include introduction 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 practical, The ESCRT Complexes: Methods and Protocols serves as a compact guide for researchers interested in establishing an integrated approach to investigate the ESCRT machinery functions in cell biology.

✦ Table of Contents

Front Matter ....Pages i-xiv
Analysis of the Archaeal ESCRT Apparatus (Rachel Y. Samson, Iain G. Duggin, Stephen D. Bell)....Pages 1-11
Genetic and Cell Biology Methods to Study ESCRTs in Drosophila melanogaster (Marco Gualtieri, Thomas Vaccari)....Pages 13-29
Functional Analysis of ESCRT-Positive Extracellular Vesicles in the Drosophila Wing Imaginal Disc (Tamás Matusek, Pascal Thérond, Maximilian Fürthauer)....Pages 31-47
Subcellular Localization of ESCRT-II in the Nematode C. elegans by Correlative Light Electron Microscopy (Céline Largeau, Emmanuel Culetto, Renaud Legouis)....Pages 49-61
Proximity Ligation Assay (PLA) to Determine the Endosomal Localization of ESCRT Subunit in Virus-Infected Cells (Binod Kumar, Mohanan Valiya Veettil, Arunava Roy, Bala Chandran)....Pages 63-72
Immuno-localization of ESCRT Proteins in Virus-Infected Cells by Fluorescence and Electron Microscopy (Keisuke Tabata, Atsuki Nara, Hiroko Omori, Eiji Morita)....Pages 73-92
Single Cell Fluorescence Ratio Image Analysis for Studying ESCRT Function in Receptor Trafficking (Jalal M. Kazan, Gergely L. Lukacs, Pirjo M. Apaja, Arnim Pause)....Pages 93-103
Genetic and Biochemical Analyses of Yeast ESCRT (Sudeep Banjade, Shaogeng Tang, Scott D. Emr)....Pages 105-116
Live Imaging of ESCRT Proteins in Microfluidically Isolated Hippocampal Axons (Veronica Birdsall, Jose C. Martinez, Lisa Randolph, Ulrich Hengst, Clarissa L. Waites)....Pages 117-128
Studying the Spatial Organization of ESCRTs in Cytokinetic Abscission Using the High-Resolution Imaging Techniques SIM and Cryo-SXT (Shai Adar-Levor, Inna Goliand, Michael Elbaum, Natalie Elia)....Pages 129-148
Three-Dimensional Surface Rendering of ESCRT Proteins Microscopy Data Using UCSF Chimera Software (Romain Le Bars, Michele W. Bianchi, Christophe Lefebvre)....Pages 149-161
Transient Expression of ESCRT Components in Arabidopsis Root Cell Suspension Culture-Derived Protoplasts (Marie-Kristin Nagel, Karin Vogel, Erika Isono)....Pages 163-174
Crystallization and Biophysical Approaches for Studying the Interactions Between the Vps4-MIT Domain and ESCRT-III Proteins (Takayuki Obita, Rieko Kojima, Mineyuki Mizuguchi)....Pages 175-187
Biochemical Approaches to Studying Caenorhabditis elegans ESCRT Functions In Vitro (Samuel Block, Amber L. Schuh, Anjon Audhya)....Pages 189-202
Purification of Recombinant ESCRT-III Proteins and Their Use in Atomic Force Microscopy and In Vitro Binding and Phosphorylation Assays (Luisa Capalbo, Ioanna Mela, Maria Alba Abad, A. Arockia Jeyaprakash, J. Michael Edwardson, Pier Paolo D’Avino)....Pages 203-217
Assessment of ESCRT Protein CHMP5 Activity on Client Protein Ubiquitination by Immunoprecipitation and Western Blotting (Francheska Son, Katharine Umphred-Wilson, Jae-Hyuck Shim, Stanley Adoro)....Pages 219-226
Purification of Plant ESCRT Proteins for Polyclonal Antibody Production (Julio Paez-Valencia, Marisa S. Otegui)....Pages 227-238
Genetic and Cytological Methods to Study ESCRT Cell Cycle Function in Fission Yeast (Imane M. Rezig, Shaun K. Bremner, Musab S. Bhutta, Ian P. Salt, Gwyn W. Gould, Christopher J. McInerny)....Pages 239-250
ESCRT Mutant Analysis and Imaging of ESCRT Components in the Model Fungus Ustilago maydis (Carl Haag, Thomas Klein, Michael Feldbrügge)....Pages 251-271
Genetic Suppressor Screen Using an Inducible FREE1-RNAi Line to Detect ESCRT Genetic Interactors in Arabidopsis thaliana (Qiong Zhao, Ying Zhu, Wenhan Cao, Jinbo Shen, Yong Cui, Shuxian Huang et al.)....Pages 273-289
Screening of Interactions with the ESCRT Machinery by a Gaussia princeps Split Luciferase-Based Complementation Assay (Rina Barouch-Bentov, Yves Jacob, Shirit Einav)....Pages 291-304
RNA Interference-Mediated Inhibition of ESCRT in Mammalian Cells (Katherine Bowers)....Pages 305-318
Back Matter ....Pages 319-322

✦ Subjects

Life Sciences; Membrane Biology; Protein Science; Cell Physiology