Gloeobacter violaceus (GLIC) Expression and Purification
5:31
GLIC Reconstitution in Asolectin Membranes
7:33
EPR Measurements
8:42
Results: Characterization of Pentameric Ligand-Gated Ion Channels
10:06
Conclusion
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The overall goal of this protocol is to demonstrate the purification, site directed spin labeling, and reconstitution of a penameric ligand-gated channel for electron paramagnetic resonance studies. Studying membrane protein dynamics in a physiolo
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This article describes methods for site-directed spin labeling and reconstitution of pentameric ligand-gated channels for Electron Paramagnetic Resonance studies. This protocol can be adapted for any membrane protein. The reconstitution method described here can also be used for patch-clamp measurements of macroscopic and single-channel currents in a defined lipid system.