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Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels
 

Site Directed Spin Labeling and EPR Spectroscopic Studies of Pentameric Ligand-Gated Ion Channels

Article DOI: 10.3791/54127-v • 11:19 min • July 4th, 2016
July 4th, 2016 •
1, 1, 1,2
1Department of Physiology and Biophysics, Case Western Reserve University, 2School of Medicine, Case Western Reserve University

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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.

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Keywords: Site Directed Spin Labeling EPR Spectroscopy Pentameric Ligand-gated Ion Channels Membrane Protein Dynamics GLIC Site-directed Mutagenesis Protein Purification Cell Lysis Membrane Protein Reconstitution
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