Liquid-Liquid Phase Coexistence and Membraneless Organelles

Liquid-Liquid Phase Coexistence and Membraneless Organelles

1st Edition - January 14, 2021

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  • Editor: Christine D. Keating
  • eBook ISBN: 9780128211601
  • Hardcover ISBN: 9780128211595

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Description

Methods in Enzymology, Volume 646, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Chapters in this new release include Methods for Studying RNA condensation/granules in vitro, RNA Dynamics in Intracellular Condensates, Methods for Viscoelastic Characterization of Liquid and Gel Condensates, Incorporating Proteins into Complex Coacervates, Methods for Study of Liquid-Liquid Phase Coexistence in Proximity to Lipid Membranes, Preparation of and Solute Partitioning in Multiphase Coacervates, Reversible photocontrol of DNA coacervation, Enzymatic Control over Coacervation, and much more.

Key Features

  • Provides the authority and expertise of leading contributors from an international board of authors
  • Presents the latest release in the Methods in Enzymology series

Readership

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

Table of Contents

  • Cover image
  • Title page
  • Table of Contents
  • Copyright
  • Contributors
  • Preface
  • Chapter One: Using a sequence-specific coarse-grained model for studying protein liquid–liquid phase separation
  • Abstract
  • 1: Introduction
  • 2: HPS model
  • 3: Simulation strategy for obtaining phase diagrams
  • 4: Summary
  • Acknowledgments
  • Chapter Two: Reconstituting bacterial cell division assemblies in crowded, phase-separated media
  • Abstract
  • 1: Introduction
  • 2: Experimental approaches and analysis
  • 3: Summary of experimental results
  • 4: Concluding remarks and future prospects
  • Acknowledgments
  • Chapter Three: Terpolymer-stabilized complex coacervates: A robust and versatile synthetic cell platform
  • Abstract
  • 1: Introduction
  • 2: Synthesis of coacervate forming materials and terpolymer
  • 3: Formation of terpolymer-stabilized coacervates
  • 4: Purification strategies
  • 5: Measurement considerations
  • 6: Conclusions and future perspectives
  • Acknowledgments
  • Chapter Four: Cell-free reconstitution of multi-condensate assemblies
  • Abstract
  • 1: Introduction
  • 2: Considerations when working with BC proteins
  • 3: Preparation of recombinant MEG-3 and PGL-3 proteins
  • 4: Fluorescent labeling of protein and RNA
  • 5: Condensation assays
  • 6: Imaging and data analysis
  • 7: Concluding remarks
  • Chapter Five: Practical considerations for generation of multi-compartment complex coacervates
  • Abstract
  • 1: Introduction
  • 2: Procedure
  • Acknowledgments
  • Chapter Six: Methods for characterizing the material properties of biomolecular condensates
  • Abstract
  • 1: Introduction
  • 2: Measuring mesoscale physical properties of liquid droplets using optical trap-induced droplet coalescence: Concepts
  • 3: Measuring mesoscale physical properties of liquid droplets using optical trap-induced droplet coalescence: Methods
  • 4: Determining nanoscale biomolecular diffusion within liquid droplets using fluorescence recovery after photobleaching (FRAP): Concepts
  • 5: Determining diffusion dynamics within liquid droplets using fluorescence recovery after photobleaching: Methods
  • 6: Single particle tracking (SPT)
  • 7: Advanced methods
  • 8: Conclusion
  • Acknowledgments
  • Chapter Seven: Small-angle X-ray scattering experiments of monodisperse intrinsically disordered protein samples close to the solubility limit
  • Abstract
  • 1: Introduction
  • 2: Resolution of structural information from SAXS
  • 3: Complications of SAXS measurements of self-assembling proteins
  • 4: IDR sample measurement in SEC-SAXS mode
  • 5: Primary data analysis
  • 6: Synchrotron SAXS beamline hardware
  • 7: Model-dependent analysis
  • 8: Implementation of SAXS measurements of single chain IDR behavior for characterizing phase behavior
  • 9: Conclusions
  • Acknowledgments
  • Chapter Eight: SAXS methods for investigating macromolecular and self-assembled polyelectrolyte complexes
  • Abstract
  • 1: Introduction
  • 2: Macromolecular polyelectrolyte complexes (PECs)
  • 3: Self-assembled PEC micelles
  • 4: Self-assembled PEC hydrogels
  • 5: Summary
  • Acknowledgments
  • Chapter Nine: Ultra-small angle neutron scattering to study droplet formation in polyelectrolyte complex coacervates
  • Abstract
  • 1: Introduction
  • 2: Model polyelectrolyte complex coacervate and liquid-liquid phase separation
  • 3: USANS instrument and experimental practice
  • 4: Related techniques
  • 5: Safety considerations and standards
  • 6: Pros and cons
  • 7: Summary
  • Acknowledgments
  • Chapter Ten: Incorporation of proteins into complex coacervates
  • Abstract
  • 1: Introduction
  • 2: Materials, equipment, and reagents
  • 3: Safety considerations
  • 4: Protocols
  • 5: Analysis and statistics
  • 6: Summary
  • Acknowledgments
  • Chapter Eleven: Measuring the activity and structure of functional RNAs inside compartments formed by liquid-liquid phase separation
  • Abstract
  • 1: Rationale
  • 2: Preparation of reagents
  • 3: RNA structure and function inside complex coacervates
  • 4: Conclusion
  • Acknowledgements
  • Chapter Twelve: Reversible photocontrol of DNA coacervation
  • Abstract
  • 1: Introduction
  • 2: Photoswitchable coacervation: The azoTAB/dsDNA system
  • 3: Synthesis and photophysical characterization of azoTAB
  • 4: Photoswitchable DNA coacervation
  • 5: Conclusions
  • Acknowledgments
  • Chapter Thirteen: Enzymatic control over coacervation
  • Abstract
  • 1: Introduction
  • 2: Enzymatic reactions in two phases
  • 3: Control over coacervation by chain length and charge density
  • 4: Protocols
  • 5: Analysis
  • 6: Summary and conclusion

Product details

  • No. of pages: 406
  • Language: English
  • Copyright: © Academic Press 2021
  • Published: January 14, 2021
  • Imprint: Academic Press
  • eBook ISBN: 9780128211601
  • Hardcover ISBN: 9780128211595

About the Serial Volume Editor

Christine D. Keating

Christine Keating is at Penn State University, PA, USA

Affiliations and Expertise

Penn State University, PA, USA

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