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 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.
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Table of Contents
PrefaceDaniel L. Minor Jr. and Henry M. Colecraft
Methods to produce and purify ion channels
1. Effective production of oligomeric membrane proteins by EarlyBac
Hiro Furukawa
2. Preparation of the Transient Receptor Potential Vanilloid 2 (TRPV2) channel for structural studies
Lejla Zubcevic
3. Production and Purification of TRPV2 and TRPV5 for Cryo-Electron Microscopy Studies
Vera Moiseekova-Bell
4. Sample Preparation of the Human TRPA1 Ion Channel for Cryo-EM Studies
Seok-Yong Lee
5. Expression and Purification of the Cardiac Sodium Channel NaV1.5 for Cryo-EM Structure Determination
William A. Catterall
6. Structural determination of human Nav1.4 and Nav1.7 using single particle cryo-electron microscopy
Nieng Yan
7. Production and Purification of ATP-Sensitive Potassium Channel Particles for Cryo-Electron Microscopy
Show-Ling Shyng
8. Production of K2P2.1 (TREK-1) for structural studies
Daniel L. Minor Jr.
9. Purification of a native nicotinic receptor
Ryan Hibbs
10. SARS-CoV-2 3a expression, purification, and reconstitution into lipid nanodiscs
Stephen Brohawn
Optical methods for probing and controlling channel function
11. Studying Ion Channel Conformation Dynamics by Encoding Coumarin as Unnatural Amino Acid
Sebastian E. Brauchi
12. Fluorescence lifetime measurement of absolute membrane potential
Evan Miller
13. Fluorescent Toxins as Ion Channel Activity Sensors
Jon Sack
14. Probing Ion Channel Macromolecular Interactions using Fluorescence Resonance Energy Transfer
Manu Ben-Johny
15. Manipulation of Ion Channel Gating with Photoswitchable Tweezers
Thomas Grutter
16. Counting Subunits in Kv Channel Complexes
Rikard Blunck
