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Bicelle samples for solid-state NMR of membrane proteins

Nature Protocols volume 2, pages 2332–2338 (2007)Cite this article

Abstract

Magnetically aligned bicelles are an excellent medium for structure determination of isotopically labeled membrane proteins by solid-state NMR spectroscopy. Bicelles are a mixture of long- and short-chain phospholipids that form bilayers in an aqueous medium and align spontaneously in a high magnetic field, for example that of an NMR spectrometer with a 1H resonance frequency between 400 and 900 MHz. Importantly, membrane proteins have been shown to be fully functional in these fully hydrated, planar bilayers under physiological conditions of pH and temperature. We describe a protocol for preparing stable protein-containing bicelles samples that yield high-resolution solid-state NMR spectra. Depending on the details of the protein and its behavior in the lipids, the time for sample preparation can vary from a few hours to several days.

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Figure 1: Schematic representation of aligned bicelle membrane.
Figure 2: One-dimensional NMR spectra of bicelle samples of membrane proteins.

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Acknowledgements

These studies were supported by research grants RO1EB 001966, RO1GM075877 and RO1EB005161, and utilized the Resource for NMR Molecular Imaging of Proteins, supported by grant P41EB002031 from the National Institute for Biomedical Imaging and Bioengineering. A.A.D.A. was supported by postdoctoral fellowship F32GM65833.

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  1. Department of Chemistry and Biochemistry, University of California at San Diego, 9500 Gilman Drive, La Jolla, 92037, California, USA

    Anna A De Angelis & Stanley J Opella

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  1. Anna A De Angelis
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  2. Stanley J Opella
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Correspondence to Stanley J Opella.

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De Angelis, A., Opella, S. Bicelle samples for solid-state NMR of membrane proteins. Nat Protoc 2, 2332–2338 (2007). https://doi.org/10.1038/nprot.2007.329

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