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MicroED Structures from Micrometer Thick Protein Crystals

Michael W. Martynowycz, Calina Glynn, Jennifer Miao, M. Jason de la Cruz, Johan Hattne, Dan Shi, Duilio Cascio, Jose Rodriguez, Tamir Gonen
doi: https://doi.org/10.1101/152504
Michael W. Martynowycz
1 Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA
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Calina Glynn
2 Department of Chemistry and Biochemistry; UCLA-DOE Institute; University of California, Los Angeles; Los Angeles, California 90095-1570, USA
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Jennifer Miao
2 Department of Chemistry and Biochemistry; UCLA-DOE Institute; University of California, Los Angeles; Los Angeles, California 90095-1570, USA
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M. Jason de la Cruz
1 Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA
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Johan Hattne
1 Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA
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Dan Shi
1 Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA
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Duilio Cascio
3 Department of Biological Chemistry, UCLA-DOE Institute; University of California, Los Angeles; Los Angeles, California 90095-1570, USA
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Jose Rodriguez
2 Department of Chemistry and Biochemistry; UCLA-DOE Institute; University of California, Los Angeles; Los Angeles, California 90095-1570, USA
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Tamir Gonen
1 Janelia Research Campus, Howard Hughes Medical Institute, 19700 Helix Drive, Ashburn, Virginia 20147, USA
4 Departments of Biological Chemistry and Physiology, David Geffen Medical School, University of California, Los Angeles, California 90095-1570, USA.
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  • For correspondence: gonen@ucla.edu)
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Abstract

Theoretical calculations suggest that crystals exceeding 100 nm thickness are excluded by dynamical scattering from successful structure determination using microcrystal electron diffraction (MicroED). These calculations are at odds with experimental results where MicroED structures have been determined from significantly thicker crystals. Here we systematically evaluate the influence of thickness on the accuracy of MicroED intensities and the ability to determine structures from protein crystals one micrometer thick. To do so, we compare ab initio structures of a human prion protein segment determined from thin crystals to those determined from crystals up to one micrometer thick. We also compare molecular replacement solutions from crystals of varying thickness for a larger globular protein, proteinase K. Our results indicate that structures can be reliably determined from crystals at least an order of magnitude thicker than previously suggested by simulation, opening the possibility for an even broader range of MicroED experiments.

Summary Atomic resolution protein structures can be determined by MicroED from crystals that surpass the theoretical maximum thickness limit by an order of magnitude.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted June 20, 2017.
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MicroED Structures from Micrometer Thick Protein Crystals
Michael W. Martynowycz, Calina Glynn, Jennifer Miao, M. Jason de la Cruz, Johan Hattne, Dan Shi, Duilio Cascio, Jose Rodriguez, Tamir Gonen
bioRxiv 152504; doi: https://doi.org/10.1101/152504
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MicroED Structures from Micrometer Thick Protein Crystals
Michael W. Martynowycz, Calina Glynn, Jennifer Miao, M. Jason de la Cruz, Johan Hattne, Dan Shi, Duilio Cascio, Jose Rodriguez, Tamir Gonen
bioRxiv 152504; doi: https://doi.org/10.1101/152504

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