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Protein and Small Molecule Structure Determination by the Cryo-EM Method MicroED

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Structural Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2305))

Abstract

Microcrystal Electron Diffraction (MicroED) is the newest cryo-electron microscopy (cryo-EM) method, with over 70 protein, peptide, and several small organic molecule structures already determined. In MicroED, micro- or nanocrystalline samples in solution are deposited on electron microscopy grids and examined in a cryo-electron microscope, ideally under cryogenic conditions. Continuous rotation diffraction data are collected and then processed using conventional X-ray crystallography programs. The protocol outlined here details how to obtain and identify the nanocrystals, how to set up the microscope for screening and for MicroED data collection, and how to collect and process data to complete high-resolution structures. For well-behaving crystals with high-resolution diffraction in cryo-EM, the entire process can be achieved in less than an hour.

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Acknowledgments

We thank all members of the Gonen laboratory, current and past and all collaborators who worked with us on MicroED applications. This work was supported by the National Institutes of Health P41GM136508. The Gonen laboratory is supported by the Howard Hughes Medical Institute.

Author information

Authors and Affiliations

  1. Department of Biological Chemistry, University of California Los Angeles, Los Angeles, CA, USA

    Emma Danelius & Tamir Gonen

  2. Department of Physiology, University of California Los Angeles, Los Angeles, CA, USA

    Emma Danelius & Tamir Gonen

  3. Howard Hughes Medical Institute, University of California Los Angeles, Los Angeles, CA, USA

    Emma Danelius & Tamir Gonen

Authors
  1. Emma Danelius
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  2. Tamir Gonen
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Corresponding author

Correspondence to Tamir Gonen .

Editor information

Editors and Affiliations

  1. The Rosalind Franklin Institute, Harwell Science Campus, Didcot, UK

    Raymond J. Owens

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Danelius, E., Gonen, T. (2021). Protein and Small Molecule Structure Determination by the Cryo-EM Method MicroED. In: Owens, R.J. (eds) Structural Proteomics. Methods in Molecular Biology, vol 2305. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1406-8_16

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  • DOI: https://doi.org/10.1007/978-1-0716-1406-8_16

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1405-1

  • Online ISBN: 978-1-0716-1406-8

  • eBook Packages: Springer Protocols

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