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EPR Techniques to Probe Insertion and Conformation of Spin-Labeled Proteins in Lipid Bilayers

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Lipid-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 974))

Abstract

Electron paramagnetic resonance (EPR) spectroscopy of spin-labeled membrane proteins is a valuable biophysical technique to study structural details and conformational transitions of proteins close to their physiological environment, e.g., in liposomes, membrane bilayers, and nanodiscs. Unlike in nuclear magnetic resonance spectroscopy, having only one or few specific side chains labeled at a time with paramagnetic probes makes the size of the object under investigation irrelevant in terms of technique sensitivity. As a drawback, extensive site-directed mutagenesis is required in order to analyze the properties of the protein under investigation. EPR can provide detailed information on side chain dynamics of large membrane proteins or protein complexes embedded in membranes with an exquisite sensitivity for flexible regions and on water accessibility profiles across the membrane bilayer. Moreover, distances between the two spin-labeled side chains in membrane proteins can be detected with high precision in the 1.5–6 nm range at cryogenic temperatures. The application of EPR to membrane proteins still presents some challenges in terms of sample preparation, sensitivity, and data interpretation; thus no ready-to-go methodological recipes can be given. However this chapter describes the state of the art in the application of nitroxide-based site-directed spin labeling EPR to membrane proteins, with specific focus on the different types of information which can be obtained with continuous wave and pulsed techniques and on the challenges in sample preparation and data analysis for functional and structural membrane protein studies.

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Authors and Affiliations

  1. Laboratory for Physical Chemistry, ETH Zurich, Zurich, Switzerland

    Enrica Bordignon & Yevhen Polyhach

Authors
  1. Enrica Bordignon
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  2. Yevhen Polyhach
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Correspondence to Enrica Bordignon .

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Editors and Affiliations

  1. Institut für Biologie, Abteilung Biophysik, Universität Kassel, Heinrich-Plett-Straße 40, Kassel, 34132, Germany

    Jörg H. Kleinschmidt

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Bordignon, E., Polyhach, Y. (2013). EPR Techniques to Probe Insertion and Conformation of Spin-Labeled Proteins in Lipid Bilayers. In: Kleinschmidt, J. (eds) Lipid-Protein Interactions. Methods in Molecular Biology, vol 974. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-275-9_15

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  • DOI: https://doi.org/10.1007/978-1-62703-275-9_15

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-274-2

  • Online ISBN: 978-1-62703-275-9

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