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The ESCRT Complexes

Methods and Protocols

  • Book
  • © 2019

Overview

Editors:
  1. Emmanuel Culetto

    1. Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ.Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette, France

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  2. Renaud Legouis

    1. Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ.Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette, France

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

Part of the book series: Methods in Molecular Biology (MIMB, volume 1998)

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Table of contents (23 protocols)

  1. Front Matter

    Pages i-xiv
    Download chapter PDF

  2. Analysis of the Archaeal ESCRT Apparatus

    • Rachel Y. Samson, Iain G. Duggin, Stephen D. Bell
    Pages 1-11

  3. Genetic and Cell Biology Methods to Study ESCRTs in Drosophila melanogaster

    • Marco Gualtieri, Thomas Vaccari
    Pages 13-29

  4. Functional Analysis of ESCRT-Positive Extracellular Vesicles in the Drosophila Wing Imaginal Disc

    • Tamás Matusek, Pascal Thérond, Maximilian Fürthauer
    Pages 31-47

  5. Subcellular Localization of ESCRT-II in the Nematode C. elegans by Correlative Light Electron Microscopy

    • Céline Largeau, Emmanuel Culetto, Renaud Legouis
    Pages 49-61

  6. 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

  7. 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

  8. 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

  9. Genetic and Biochemical Analyses of Yeast ESCRT

    • Sudeep Banjade, Shaogeng Tang, Scott D. Emr
    Pages 105-116

  10. 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

  11. 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

  12. Three-Dimensional Surface Rendering of ESCRT Proteins Microscopy Data Using UCSF Chimera Software

    • Romain Le Bars, Michele W. Bianchi, Christophe Lefebvre
    Pages 149-161

  13. Transient Expression of ESCRT Components in Arabidopsis Root Cell Suspension Culture-Derived Protoplasts

    • Marie-Kristin Nagel, Karin Vogel, Erika Isono
    Pages 163-174

  14. 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

  15. Biochemical Approaches to Studying Caenorhabditis elegans ESCRT Functions In Vitro

    • Samuel Block, Amber L. Schuh, Anjon Audhya
    Pages 189-202

  16. 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

  17. 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

  18. Purification of Plant ESCRT Proteins for Polyclonal Antibody Production

    • Julio Paez-Valencia, Marisa S. Otegui
    Pages 227-238

  19. 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

  20. ESCRT Mutant Analysis and Imaging of ESCRT Components in the Model Fungus Ustilago maydis

    • Carl Haag, Thomas Klein, Michael Feldbrügge
    Pages 251-271
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Keywords

About this book

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.

Editors and Affiliations

  • Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Univ.Paris-Sud, Université Paris-Saclay, Gif-sur-Yvette, France

    Emmanuel Culetto, Renaud Legouis

Bibliographic Information

  • Book Title: The ESCRT Complexes

  • Book Subtitle: Methods and Protocols

  • Editors: Emmanuel Culetto, Renaud Legouis

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-9492-2

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

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

  • Hardcover ISBN: 978-1-4939-9491-5Published: 02 July 2019

  • Softcover ISBN: 978-1-4939-9494-6Published: 14 August 2020

  • eBook ISBN: 978-1-4939-9492-2Published: 27 June 2019

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIV, 322

  • Number of Illustrations: 29 b/w illustrations, 43 illustrations in colour

  • Topics: Membrane Biology, Protein Science, Cell Physiology

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