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PELDOR/DEER: An Electron Paramagnetic Resonance Method to Study Membrane Proteins in Lipid Bilayers

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Biophysics of Membrane Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2168))

Abstract

Every membrane protein is involved in close interactions with the lipid environment of cellular membranes. The annular lipids, that are in direct contact with the polypeptide, can in principle be seen as an integral part of its structure, akin to the first hydration shell of soluble proteins. It is therefore desirable to investigate the structure of membrane proteins and especially their conformational flexibility under conditions that are as close as possible to their native state. This can be achieved by reconstituting the protein into proteoliposomes, nanodiscs, or bicelles. In recent years, PELDOR/DEER spectroscopy has proved to be a very useful method to study the structure and function of membrane proteins in such artificial membrane environments. The technique complements both X-ray crystallography and cryo-EM and can be used in combination with virtually any artificial membrane environment and under certain circumstances even in native membranes. Of the above-mentioned membrane mimics, bicelles are currently the least often used for PELDOR studies, although they offer some advantages, especially their ease of use. Here, we provide a step-by-step protocol for studying a bicelle reconstituted membrane protein with PELDOR/DEER spectroscopy.

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Acknowledgments

We thank our colleague Nicole Florin for technical assistance. Ki.B. is supported by a Biotechnology and Biological Sciences Research Council (BBSRC) Doctoral Training Partnership (DTP). This work was supported by the Medical Research Council (MR/N020103/1 to K.B.). M.F. Peter is supported by a fellowship of the Konrad-Adenauer-Stiftung.

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Authors and Affiliations

  1. Institute of Structural Biology, Biomedical Center, University of Bonn, Bonn, Germany

    Martin F. Peter & Gregor Hagelueken

  2. Department of Life Sciences, Imperial College London, London, UK

    Kiran Bountra & Konstantinos Beis

  3. Research Complex at Harwell, Oxford, UK

    Kiran Bountra & Konstantinos Beis

Authors
  1. Martin F. Peter
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  2. Kiran Bountra
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  3. Konstantinos Beis
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  4. Gregor Hagelueken
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Correspondence to Gregor Hagelueken .

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Editors and Affiliations

  1. School of Clinical and Applied Sciences, Leeds Beckett University, Leeds, UK

    Vincent L. G. Postis

  2. Astbury Centre for Structural Molecular Biology, School of Biomedical Sciences; Molecular and Integrative Biosciences, Faculty of Biological and Environmental Sciences, University of Leeds, Leeds, UK; University of Helsinki, Helsinki, Finland

    Adrian Goldman

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Peter, M.F., Bountra, K., Beis, K., Hagelueken, G. (2020). PELDOR/DEER: An Electron Paramagnetic Resonance Method to Study Membrane Proteins in Lipid Bilayers. In: Postis, V.L.G., Goldman, A. (eds) Biophysics of Membrane Proteins. Methods in Molecular Biology, vol 2168. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0724-4_15

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  • DOI: https://doi.org/10.1007/978-1-0716-0724-4_15

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