Ion Channels: Channel Biochemistry, Reconstitution, and Function
- 1st Edition, Volume 652 - May 29, 2021
- Imprint: Academic Press
- Editors: Daniel L. Minor, Henry M. Colecraft
- Language: English
- Hardback ISBN:9 7 8 - 0 - 3 2 3 - 8 5 3 7 4 - 3
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 8 5 3 7 5 - 0
Ion Channels Part A, Volume 651 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a variety of new… Read more
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Request a sales quoteIon Channels Part A, Volume 651 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on a variety of new developments on the topic. Each chapter is written by an international board of authors.
- Provides the authority and expertise of leading contributors from an international board of authors
- Presents the latest release in the Methods in Enzymology series
Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists
- Cover image
- Title page
- Table of Contents
- Copyright
- Contributors
- Preface
- Section I: Structure/function relationships
- Chapter One: Correlating ion channel structure and function
- Abstract
- 1: Introduction
- 2: Nanodiscs
- 3: Liposome based assays
- 4: Examples of studies correlating structure and function of ion channels
- 5: Protocols
- 6: Conclusion
- Acknowledgments
- Chapter Two: TRPV3 expression and purification for structure determination by Cryo-EM
- Abstract
- 1: Introduction
- 2: Key resources table
- 3: Expression, purification, and cryo-EM of TRPV3
- 4: Summary
- Chapter Three: Methods to study phosphoinositide regulation of ion channels
- Abstract
- 1: Introduction
- 2: Applying phosphoinositides directly in excised patches or intact cells
- 3: Manipulating endogenous phosphoinositides in intact cells
- 4: Monitoring phosphoinositides
- 5: Techniques using purified proteins
- 6: Summary
- Acknowledgments
- Chapter Four: Recombinant expression and purification of pentameric ligand-gated ion channels for Cryo-EM structural studies
- Abstract
- 1: Introduction
- 2: Key resources table
- 3: Materials and equipment
- 4: Cloning of 5-HT3AR and GlyRα1 in pFastBac1 vector
- 5: Large-scale expression and purification of 5-HT3AR
- 6: Sample preparation for 5-HT3AR cryo-EM imaging
- 7: Large-scale expression and purification of GlyRα1
- 8: Reconstitution of pLGICs in lipid nanodisc
- 9: Sample preparation for GlyRα1 cryo-EM imaging
- 10: Summary
- Acknowledgments
- Chapter Five: Detection of ligand binding to purified HCN channels using fluorescence-based size exclusion chromatography
- Abstract
- 1: Introduction
- 2: Relevant issues
- 3: Equipment
- 4: Chemicals
- 5: Protocol
- 6: Summary
- Section II: Structural and chemical methods to assess channel dynamics
- Chapter Six: Structural dynamics of channels and transporters by high-speed atomic force microscopy
- Abstract
- 1: Introduction
- 2: Concepts
- 3: Equipment and chemicals
- 4: Protocols
- 5: Summary
- Chapter Seven: Crosslinking glutamate receptor ion channels
- Abstract
- 1: Introduction
- 2: Receptor crosslinking with introduced disulfide bonds
- 3: Photocrosslinking with genetically-encoded unnatural amino acids
- 4: Fast perfusion coupled to chemical modification and state-dependent crosslinking
- 5: Biochemistry: Photoactivated Crosslinkers and purification of GluA2
- 6: Modeling of the effects of crosslinking
- 7: General limitations
- 8: Summary and perspectives
- Acknowledgments
- Chapter Eight: Single molecule FRET methodology for investigating glutamate receptors
- Abstract
- 1: Background
- 2: Experimental design for the application of smFRET on iGluRs
- 3: Before you begin
- 4: Materials and equipment
- 5: Key resources table
- 6: Step-by-step method details
- 7: Quantification and statistical analysis
- 8: Advantages
- 9: Limitations
- 10: Optimization and troubleshooting
- 11: Summary
- Acknowledgments
- Chapter Nine: Interrogating permeation and gating of Orai channels using chemical modification of cysteine residues
- Abstract
- 1: Introduction
- 2: Substituted cysteine accessibility analysis (SCAM)
- 3: SCAM analysis of the STIM1-gated human Orai1 channel
- 4: Materials, equipment and reagents
- 5: Detailed steps
- 6: Analysis and statistics
- 7: Troubleshooting and optimization
- 8: Conclusions
- Acknowledgment
- Section III: Channel reconstitution and studies in bilayers
- Chapter Ten: A quantitative flux assay for the study of reconstituted Cl− channels and transporters
- Abstract
- 1: Introduction
- 2: Protocol
- 3: Protocol variations
- 4: Discussion
- Chapter Eleven: Ion channel reconstitution in lipid bilayers
- Abstract
- 1: Introduction
- 2: Advantages of lipid bilayer studies
- 3: Disadvantages of lipid bilayer
- 4: Methods
- 5: Analysis and statistics
- 6: Alternative methods/procedures
- 7: Troubleshooting and optimization
- 8: Summary
- Chapter Twelve: Combining in vitro translation with nanodisc technology and functional reconstitution of channels in planar lipid bilayers
- Abstract
- 1: Introduction
- 2: Cell-free protein synthesis into nanodiscs: A new entry into functional bilayer reconstitution
- 3: Combining in vitro translation into nanodiscs and functional reconstitution
- 4: Protocols and handling
- 5: Conclusion
- Acknowledgments
- Section IV: Methods to assess oligomerization
- Chapter Thirteen: The application of Poisson distribution statistics in ion channel reconstitution to determine oligomeric architecture
- Abstract
- 1: Introduction
- 2: The Poisson distribution in theory and in experiment
- 3: Experimental design: Measurement of f0
- 4: Performing the assay: A case study of fluoride channel oligomeric architecture
- 5: Summary
- Acknowledgment
- Edition: 1
- Volume: 652
- Published: May 29, 2021
- No. of pages (Hardback): 356
- No. of pages (eBook): 356
- Imprint: Academic Press
- Language: English
- Hardback ISBN: 9780323853743
- eBook ISBN: 9780323853750
DM
Daniel L. Minor
Daniel L. Minor, Professor, Cardiovascular Research Institute, University of California San Francisco, San Francisco, CA, USA
Affiliations and expertise
Professor, University of California San Francisco, CA, USAHC
Henry M. Colecraft
Henry M. Colecraft, John C. Dalton Professor of Physiology and Cellular Biophysics & Cellular Biophysics, Columbia University College of Physicians & Surgeons, New York, NY, USA
Affiliations and expertise
Professor of Physiology and Cellular Biophysics and Cellular Biophysics, Columbia University College of Physicians and Surgeons, NY, USARead Ion Channels: Channel Biochemistry, Reconstitution, and Function on ScienceDirect