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Amphipols: A General Introduction and Some Protocols

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Membrane Proteins Production for Structural Analysis

Abstract

Membrane proteins (MPs) exhibit a broad range of activities, which are crucial for cell survival. They can be pumps, channels, enzymes, scaffolds, signal transmitters, or a combination of these functions. Understanding their molecular mechanisms generally requires their extraction out of membranes and their purification. Solubilization and isolation are usually carried out using detergents, which disrupt the membrane and adsorb onto the hydrophobic surface of the transmembrane domain of MPs, keeping them water soluble. Detergents, however, tend to inactivate most MPs more or less rapidly, making their biochemical and biophysical studies challenging. Specially designed amphipathic polymers called “amphipols” (APols) have been developed with the view of improving the stability of MPs in aqueous solutions. In this chapter, the properties of APols and of the complexes they form with MPs are summarized, and a brief overview of APol applications that have been validated thus far is presented. Five experimental protocols are described in detail: (1) trapping MPs in APols, (2) measuring the amount of APol bound per MP, (3) APol-assisted folding of MPs, (4) APol-assisted production of MPs by cell-free expression, and (5) immobilizing MPs onto solid surfaces for screening purposes using functionalized APols.

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Acknowledgments

We would like to thank L. J. Catoire for reading the protocol of APol-assisted MP folding. The development of amphipols has been mainly supported by the Centre National de la Recherche Scientifique, the Human Frontier Science Program Organization (RG00223/2000-M), and the European Community (BIO4-CT98-0269 and STREP LSHG-CT-2005-513770 Innovative Tools for Membrane Protein Structural Proteomics).

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Authors and Affiliations

  1. Laboratory of Physico-Chemical Biology of Membrane Proteins, UMR-CNRS 7099, Institute of Physico-Chemical Biology, and Université Paris Diderot, 13, rue Pierre-et-Marie-Curie, 75005, Paris, France

    Manuela Zoonens, Francesca Zito & Jean-Luc Popot

  2. Bio-Nanotechnology Laboratory, Department of Neuroscience and Pharmacology & Nano-Science Center, University of Copenhagen, Universitetsparken 5, 2100, Copenhagen, Denmark

    Karen L. Martinez

Authors
  1. Manuela Zoonens
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  2. Francesca Zito
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  3. Karen L. Martinez
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  4. Jean-Luc Popot
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Corresponding author

Correspondence to Manuela Zoonens .

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Editors and Affiliations

  1. UMR-CNRS 7275, Institute of Molecular and Cellular Pharmacology, Valbonne, France

    Isabelle Mus-Veteau

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Zoonens, M., Zito, F., Martinez, K., Popot, JL. (2014). Amphipols: A General Introduction and Some Protocols. In: Mus-Veteau, I. (eds) Membrane Proteins Production for Structural Analysis. Springer, New York, NY. https://doi.org/10.1007/978-1-4939-0662-8_7

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