Chapter title |
Random Spherically Constrained Single-Particle (RSC) Method to Study Voltage-Gated Ion Channels
|
---|---|
Chapter number | 20 |
Book title |
Potassium Channels
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7362-0_20 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7361-3, 978-1-4939-7362-0
|
Authors |
Liguo Wang |
Abstract |
To study membrane protein structures using cryo-Electron Microscopy (cryo-EM), membrane proteins are usually extracted from cell membranes and solubilized in detergents. To restore the lipid bilayer environment of membrane proteins, a method called "random spherically constrained" (RSC) single-particle cryo-EM has been developed. The RSC platform establishes the lipid environment for membrane proteins and makes it possible, for the first time, to apply the desired transmembrane potential to trap voltage-gated ion channels in the desired functional states (e.g., deactivated voltage sensor at -120 mV) for structural analysis. No rupture or leakage was observed during the establishment of the transmembrane potential. The spherical geometry of liposomes is used as a constraint to accurately determine the orientation of the inserted membrane protein. |
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