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Phasing Electron Diffraction Data by Molecular Replacement: Strategy for Structure Determination and Refinement

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Electron Crystallography of Soluble and Membrane Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 955))

Abstract

Electron crystallography is arguably the only electron cryomicroscopy (cryo EM) technique able to deliver atomic resolution data (better then 3 Å) for membrane proteins embedded in a membrane. The progress in hardware improvements and sample preparation for diffraction analysis resulted in a number of recent examples where increasingly higher resolutions were achieved. Other chapters in this book detail the improvements in hardware and delve into the intricate art of sample preparation for microscopy and electron diffraction data collection and processing. In this chapter, we describe in detail the protocols for molecular replacement for electron diffraction studies. The use of a search model for phasing electron diffraction data essentially eliminates the need of acquiring image data rendering it immune to aberrations from drift and charging effects that effectively lower the attainable resolution.

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Acknowledgments

Research in the Gonen laboratory is supported by the American Diabetes Association Award # 1-09-CD-05 and by the National Institutes of Health R01GM079233 and U54GM094598 as well as the Howard Hughes Medical Institute.

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Authors and Affiliations

  1. Janelia Farm Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA

    Goragot Wisedchaisri & Tamir Gonen

Authors
  1. Goragot Wisedchaisri
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  2. Tamir Gonen
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Corresponding author

Correspondence to Tamir Gonen .

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Editors and Affiliations

  1. School of Biology, Georgia Institute of Technology, Ferst Dr. 310, Atlanta, 30332, Georgia, USA

    Ingeborg Schmidt-Krey

  2. , Dept. Biochemistry & Biophysics, University of California, San Francisco, 16th St. 600, San Francisco, 94158, California, USA

    Yifan Cheng

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Wisedchaisri, G., Gonen, T. (2013). Phasing Electron Diffraction Data by Molecular Replacement: Strategy for Structure Determination and Refinement. In: Schmidt-Krey, I., Cheng, Y. (eds) Electron Crystallography of Soluble and Membrane Proteins. Methods in Molecular Biology, vol 955. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-176-9_14

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  • DOI: https://doi.org/10.1007/978-1-62703-176-9_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-175-2

  • Online ISBN: 978-1-62703-176-9

  • eBook Packages: Springer Protocols

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