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Cryo Electron Microscopy of TRP Channels

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TRP Channels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1987))

Abstract

Cryo electron microscopy (cryo-EM) is a powerful technique that can be used to elucidate the structural architecture of a protein molecule in a physiologically relevant environment. In this method, purified protein is frozen in its aqueous buffer in a thin layer of vitreous ice in which the biological macromolecules are embedded in various orientations. Images of this frozen sample are collected with an electron microscope, and the data is processed using different software algorithms resulting in high-resolution structures of the protein. Proteins in the presence of various ligands or other macromolecular complexes can also be studied by this method. Here, we present a protocol for the purification and vitrification of TRP channels for single particle cryo-EM.

Amrita Samanta and Taylor E. T. Hughes contributed equally to this work.

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Authors and Affiliations

  1. Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Amrita Samanta, Taylor E. T. Hughes & Vera Y. Moiseenkova-Bell

Authors
  1. Amrita Samanta
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  2. Taylor E. T. Hughes
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  3. Vera Y. Moiseenkova-Bell
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Corresponding author

Correspondence to Vera Y. Moiseenkova-Bell .

Editor information

Editors and Affiliations

  1. Instituto de Biología, Molecular y Celular, Universidad Miguel Hernández, Elche, Alicante, Spain

    Antonio Ferrer-Montiel

  2. Experimental Anesthesiology and Pain Research, Department of Anesthesiology and Intensive Care Medicine, University Hospital Cologne, University of Cologne, Cologne, Germany

    Tim Hucho

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Samanta, A., Hughes, T.E.T., Moiseenkova-Bell, V.Y. (2019). Cryo Electron Microscopy of TRP Channels. In: Ferrer-Montiel, A., Hucho, T. (eds) TRP Channels. Methods in Molecular Biology, vol 1987. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9446-5_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9446-5_3

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9445-8

  • Online ISBN: 978-1-4939-9446-5

  • eBook Packages: Springer Protocols

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