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Random Spherically Constrained Single-Particle (RSC) Method to Study Voltage-Gated Ion Channels

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Potassium Channels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1684))

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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Author information

Authors and Affiliations

  1. Department of Biological Structure, University of Washington, 1959 NE Pacific St., Box 357420, Seattle, WA, 98195, USA

    Liguo Wang

Authors
  1. Liguo Wang
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Correspondence to Liguo Wang .

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Editors and Affiliations

  1. Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, Oregon, USA

    Show-Ling Shyng

  2. Department of Physiology and Pharmacology, Oregon Health and Science University, Portland, Oregon, USA

    Francis I. Valiyaveetil

  3. The Vollum Institute, Oregon Health and Science University, Portland, Oregon, USA

    Matt Whorton

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Wang, L. (2018). Random Spherically Constrained Single-Particle (RSC) Method to Study Voltage-Gated Ion Channels. In: Shyng, SL., Valiyaveetil, F., Whorton, M. (eds) Potassium Channels. Methods in Molecular Biology, vol 1684. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7362-0_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7362-0_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7361-3

  • Online ISBN: 978-1-4939-7362-0

  • eBook Packages: Springer Protocols

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