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Imaging of Transmembrane Proteins Directly Incorporated Within Supported Lipid Bilayers Using Atomic Force Microscopy

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Nanoimaging

Part of the book series: Methods in Molecular Biology ((MIMB,volume 950))

Abstract

Structural analysis of transmembrane proteins remains a challenge in biology, mainly due to their difficulty in being overexpressed and the required use of detergents that impair different steps of biochemistry classically used to obtain 3D crystals. In this context, we have developed a new technique for protein incorporation within supported lipid bilayers that only requires a few picomoles of protein per assay. Proteins are directly inserted into a detergent-destabilized bilayer that can be imaged in buffer with atomic force microscopy (AFM) allowing structural analysis down to sub-nanometer lateral resolution. In this chapter, we describe the main guidelines for this technique, from the choice of detergent to the requirements for AFM high-resolution imaging.

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Acknowledgments

The research has been supported by Institut Curie (DL), CNRS (DL and PEM), and INSERM (P.E.M.) and granted by the ANR grant “ANR-06-PCVI-0021-01, AFM -MB-PROT.” We are grateful to our collaborators involved in the project: Drs. A. Berquand, J. Hernández-Borrell, C. le Grimellec, F. Gubellini, P. Dosset, L. Picas, and B. Seantier.

Author information

Authors and Affiliations

  1. Institut Curie, Paris, France

    Daniel Levy

  2. Institut National de la Santé et de la Recherche Médicale, Montpellier, France

    Pierre-Emmanuel Milhiet

  3. Centre National de la Recherche Scientifique, Centre de Biochimie Structurale, Université de Montpellier, Montpellier, France

    Pierre-Emmanuel Milhiet

Authors
  1. Daniel Levy
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  2. Pierre-Emmanuel Milhiet
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Corresponding author

Correspondence to Pierre-Emmanuel Milhiet .

Editor information

Editors and Affiliations

  1. Nat. Inst. Biomedical Imaging & Bioengin, National Institutes of Health, South Drive 13, Bethesda, 20892, Maryland, USA

    Alioscka A. Sousa

  2. , National Cancer Institute, National Institutes of Health, Center Drive 10, Bethesda, 20892, USA

    Michael J. Kruhlak

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Levy, D., Milhiet, PE. (2013). Imaging of Transmembrane Proteins Directly Incorporated Within Supported Lipid Bilayers Using Atomic Force Microscopy. In: Sousa, A., Kruhlak, M. (eds) Nanoimaging. Methods in Molecular Biology, vol 950. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-137-0_19

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  • DOI: https://doi.org/10.1007/978-1-62703-137-0_19

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-136-3

  • Online ISBN: 978-1-62703-137-0

  • eBook Packages: Springer Protocols

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