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

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 physiologically relevant environment is crucial to fully understand how this membrane transport protein works. And because of these needs, I think site directed spin labeling and EPR technique is ideally suited.

The main advantage of this technique is that it's not limited by the size of the protein. Therefore, site directed spin labeling and EPR spectroscopy would be a technique of choice if you're wanting to study large protein dynamics or proteins reconstituted in membranes. Introduce single cysteine mutations in the GLIC gene by site directed mutagenesis.