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

Sandip Basak; Soumili Chatterjee; Sudha Chakrapani

doi:10.3791/54127

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Química

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Local Dirigido Labeling rotação e EPR espectroscópicos Estudos de Canais pentamérica Ligand-Gated Ion

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

JoVE Journal
Química

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JoVE Journal Química

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

Local Dirigido Labeling rotação e EPR espectroscópicos Estudos de Canais pentamérica Ligand-Gated Ion

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10,557 Views

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11:19 min

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July 04, 2016

DOI:

10.3791/54127-v

DOI:

10.3791/54127-v

•

11:19 min

July 04, 2016

•

4 Views

Sandip Basak¹, Soumili Chatterjee¹, Sudha Chakrapani^1,2

¹Department of Physiology and Biophysics,Case Western Reserve University, ²School of Medicine,Case Western Reserve University

Transcrição

Summary

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.