Ion Channels: Channel Production and Optical Methods
- 1st Edition, Volume 653 - June 5, 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 6 - 7
- eBook ISBN:9 7 8 - 0 - 3 2 3 - 8 5 3 7 7 - 4
Ion Channels Part B, Volume 652 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of top… Read more
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Request a sales quoteIon Channels Part B, Volume 652 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of topics, including NMDAR, Pannexin, and CALHM, Making NaV1.4 and NaV1.7, TRPVs, Purification native nAChRs, GABAR Radu Aricescu, TRPV5/2, NaV1.5, KATP, TRPA1, TREK-1, SARS-CoV-2 3a ion channel, Ion channel conformational dynamics by encoded unnatural amino acid, Fluorescence lifetime measurement of absolute membrane potential, Fluorescent Toxins as Activity Sensors, FRET Analyses of Ion Channel Protein-Protein Interactions, Control of Ion Channel Gating with Photo-Switchable Tweezers, and Counting Subunits in Kv Channel Complexes.
- 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: Methods to produce and purify ion channels
- Chapter One: Effective production of oligomeric membrane proteins by EarlyBac-insect cell system
- Abstract
- 1: Introduction
- 2: Initial observation
- 3: Screening of early promoters for optimal expression
- 4: Optimization by addition of enhancer and UTR
- 5: Usage of FITC-labeled 1D4 antibodies for ab-FSEC
- 6: Expression and purification of NMDAR and CALHM
- 7: Additional note and concluding remark
- Acknowledgment
- Chapter Two: Preparation of the Transient Receptor Potential Vanilloid 2 (TRPV2) channel for structural studies
- Abstract
- 1: Introduction
- 2: Preparation of TRPV2 for structural studies
- 3: Protocol
- 4: Alternative methods
- 5: Summary
- Acknowledgments
- Conflict of interest
- Chapter Three: Production and purification of TRPV2 and TRPV5 for structural and functional studies
- Abstract
- 1: Introduction
- 2: Methods
- 3: Summary
- 4: Materials table
- Chapter Four: Sample preparation of the human TRPA1 ion channel for cryo-EM studies
- Abstract
- 1: Introduction
- 2: Key resources table
- 3: Step-by step method details
- 4: Summary
- Acknowledgments
- Chapter Five: Expression and purification of the cardiac sodium channel NaV1.5 for cryo-EM structure determination
- Abstract
- 1: Introduction
- 2: Difficulties in structure determination of mammalian voltage-gated sodium channels
- 3: Expression and purification of NaV1.5
- 4: Protocol
- 5: Structure of NaV1.5
- 6: Summary
- Acknowledgments
- Chapter Six: Structural determination of human Nav1.4 and Nav1.7 using single particle cryo-electron microscopy
- Abstract
- 1: Introduction
- 2: Structural determination of human Nav1.4
- 3: Structural determination of human Nav1.7
- 4: Conclusions and perspectives
- Acknowledgments
- Chapter Seven: Production and purification of ATP-sensitive potassium channel particles for cryo-electron microscopy
- Abstract
- 1: Introduction
- 2: General considerations: Expression system and construct design
- 3: Protocol of KATP channel production and purificiation
- 4: Sample preparation for electron microscopy
- 5: Summary
- Acknowledgments
- Chapter Eight: Production of K2P2.1 (TREK-1) for structural studies
- Abstract
- 1: Introduction: K2P2.1 (TREK-1), a model polymodal ion channel
- 2: Expression and purification of K2P2.1 (TREK-1) using Pichia pastoris
- 3: Generation of anti-GFP nanobody Sepharose resin
- 4: Generation of 3C protease
- 5: Reconstitution of K2P2.1 (TREK-1) in nanodiscs
- 6: Summary
- Chapter Nine: Purification of a native nicotinic receptor
- Abstract
- 1: Introduction
- 2: Equipment and materials
- 3: Receptor purification and reconstitution in lipid nanodisc
- 4: Summary
- Acknowledgment
- Chapter Ten: SARS-CoV-2 3a expression, purification, and reconstitution into lipid nanodiscs
- Abstract
- 1: Introduction
- 2: Before you begin
- 3: Key resources table
- 4: Materials and equipment
- 5: Step-by-step method details
- 6: Expected outcomes
- 7: Advantages
- 8: Limitations
- 9: Safety considerations and standards
- 10: Alternative methods/procedures
- Acknowledgments
- Section II: Optical methods for probing and controlling channel function
- Chapter Eleven: Studying ion channel conformation dynamics by encoding coumarin as unnatural amino acid
- Abstract
- 1: Introduction
- 2: Materials, reagents, and equipment
- 3: Experimental procedures
- 4: Safety considerations and standards
- 5: Analysis and statistics
- 6: Conclusions
- 7: Pros and cons
- 8: Troubleshooting and optimization
- Acknowledgments
- Chapter Twelve: VoltageFluor dyes and fluorescence lifetime imaging for optical measurement of membrane potential
- Abstract
- 1: Introduction
- 2: Rationale
- 3: Protocol
- 4: Analysis
- 5: Summary
- Chapter Thirteen: Fluorescent toxins as ion channel activity sensors
- Abstract
- 1: Introduction
- 2: Conformation-sensitive ligands to image structural changes
- 3: Endogenous voltage-sensor activation probe synthesis and imaging
- 4: Protocols
- 5: Analysis
- 6: Summary
- Acknowledgments
- Chapter Fourteen: Probing ion channel macromolecular interactions using fluorescence resonance energy transfer
- Abstract
- 1: Introduction
- 2: Obtaining FRET efficiency from live cells
- 3: General overview of three-cube fret measurements
- 4: Deducing stoichiometry and relative binding affinities from FRET 2-hybrid assays
- 5: Protocol
- 6: Applications
- 7: Challenges and troubleshooting
- 8: Summary
- Acknowledgment
- Chapter Fifteen: Manipulation of ion channel gating with photoswitchable tweezers
- Abstract
- 1: Introduction
- 2: Strategic design
- 3: Manipulation of P2X2 ion channel activity with photoswitchable tweezers
- 4: Method
- 5: Analysis and interpretation
- 6: Summary
- Chapter Sixteen: Determining stoichiometry of ion channel complexes using single subunit counting
- Abstract
- 1: Introduction
- 2: Specific labeling of membrane proteins
- 3: Expression control and expression system
- 4: Analysis of single subunit counting
- 5: Limitations
- 6: Alternative methods
- 7: Protocol
- Edition: 1
- Volume: 653
- Published: June 5, 2021
- No. of pages (Hardback): 420
- No. of pages (eBook): 420
- Imprint: Academic Press
- Language: English
- Hardback ISBN: 9780323853767
- eBook ISBN: 9780323853774
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 Production and Optical Methods on ScienceDirect