Abstract
The structure determination of membrane protein in lipid environment can be carried out using cryo-electron microscopy combined with the recent development of data collection and image processing. We describe a protocol to study assemblies or stacks of membrane protein reconstituted into a lipid membrane using both cryo-electron tomography and single particle analysis, which is an alternative approach to electron crystallography for solving 3D structure. We show the organization of the successive layers of OprM molecules revealing the protein–protein interactions between OprM molecules of two successive lipid bilayers.
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Acknowledgments
We would like to thank Dr. Achilleas Frangakis for fruitful discussions and for giving us access to the EM facilities of EMBL (Heidelberg) and Pr. A. Ducruix for providing purified membrane protein samples. S. Trépout is recipient of PhD fellowship from French Ministry of Education and Research and Technology (MENRT). This work has been supported in part by Agence Nationale de la Recherche (ANR-06-PCV1-001) and by Conseil Régional d’Aquitaine (20071302007) grants.
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Trépout, S., Taveau, JC., Lambert, O. (2010). Structure Determination of Membrane Protein by Both Cryo-Electron Tomography and Single Particle Analysis. In: Lacapère, JJ. (eds) Membrane Protein Structure Determination. Methods in Molecular Biology, vol 654. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-762-4_11
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DOI: https://doi.org/10.1007/978-1-60761-762-4_11
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