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Single-Particle Cryo-EM of Membrane Proteins

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Book cover Structure and Function of Membrane Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2302))

Abstract

In the recent years, the protein databank has been fueled by the exponential growth of high-resolution electron cryo-microscopy (cryo-EM) structures. This trend will be further accelerated through the continuous software and method developments and the increasing availability of imaging centers, which will open cryo-EM to a wide array of researchers with their diverse scientific goals and questions. Especially for structural biology of membrane proteins, cryo-EM offers significant advantages as it can overcome multiple limitations of classical methods. Most importantly, in cryo-EM, the sample is prepared as a vitrified suspension, which abolishes the need for crystallization, reduces the required sample amount and allows usage of a wide arsenal of hydrophobic environments. Despite recent improvements, high-resolution cryo-EM still poses some significant challenges, and standardized procedures, especially for the characterization of membrane proteins, are missing. While there can be no ultimate recipe toward a high-resolution cryo-EM structure for every membrane protein, certain factors seem to be universally relevant. Here, we share the protocols that have been successfully used in our laboratory. We hope that this may be a useful resource to other researchers in the field and may increase their chances of success.

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Acknowledgments

We are grateful for funding through the DFG Mo2752/2, the Cluster of Excellence Frankfurt EXC 115, as well as support from the Max Planck Society. We would like to thank Deryck Mills and Prof. Werner Kühlbrandt for their support and access to the outstanding cryo-EM facility.

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Authors and Affiliations

  1. Max-Planck Institute of Biophysics, Frankfurt, Germany

    Dovile Januliene & Arne Moeller

  2. Department of Structural Biology, University of Osnabrück, Osnabrück, Germany

    Dovile Januliene & Arne Moeller

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  1. Dovile Januliene
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  2. Arne Moeller
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Correspondence to Arne Moeller .

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Editors and Affiliations

  1. School of Biological Sciences and School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA, USA

    Ingeborg Schmidt-Krey

  2. School of Physics, Georgia Institute of Technology, Atlanta, GA, USA

    James C. Gumbart

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Januliene, D., Moeller, A. (2021). Single-Particle Cryo-EM of Membrane Proteins. In: Schmidt-Krey, I., Gumbart, J.C. (eds) Structure and Function of Membrane Proteins. Methods in Molecular Biology, vol 2302. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1394-8_9

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