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Title: Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED

Crystallographic analysis of macromolecules depends on large, well-ordered crystals, which often require significant effort to obtain. Even sizable crystals sometimes suffer from pathologies that render them inappropriate for high-resolution structure determination. Here we show that fragmentation of large, imperfect crystals can provide a simple path for high-resolution structure determination by serial femtosecond crystallography or the cryoEM method MicroED.
Authors:
ORCiD logo [1] ; ORCiD logo [1] ;  [1] ;  [2] ;  [2] ;  [1] ;  [2] ;  [2] ;  [3] ;  [3] ;  [3] ;  [3] ;  [3] ;  [2] ;  [1]
  1. Howard Hughes Medical Inst., Ashburn, VA (United States). Janelia Research Campus
  2. Univ. of California, Los Angeles, CA (United States)
  3. Univ. of Pittsburgh, PA (United States)
Publication Date:
Grant/Contract Number:
FC02-02ER63421
Type:
Accepted Manuscript
Journal Name:
Nature Methods
Additional Journal Information:
Journal Volume: 14; Journal Issue: 4; Journal ID: ISSN 1548-7091
Publisher:
Nature Publishing Group
Research Org:
Univ. of California, Los Angeles, CA (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1463137

de la Cruz, M. Jason, Hattne, Johan, Shi, Dan, Seidler, Paul, Rodriguez, Jose, Reyes, Francis E., Sawaya, Michael R., Cascio, Duilio, Weiss, Simon C., Kim, Sun Kyung, Hinck, Cynthia S., Hinck, Andrew P., Calero, Guillermo, Eisenberg, David, and Gonen, Tamir. Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED. United States: N. p., Web. doi:10.1038/nmeth.4178.
de la Cruz, M. Jason, Hattne, Johan, Shi, Dan, Seidler, Paul, Rodriguez, Jose, Reyes, Francis E., Sawaya, Michael R., Cascio, Duilio, Weiss, Simon C., Kim, Sun Kyung, Hinck, Cynthia S., Hinck, Andrew P., Calero, Guillermo, Eisenberg, David, & Gonen, Tamir. Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED. United States. doi:10.1038/nmeth.4178.
de la Cruz, M. Jason, Hattne, Johan, Shi, Dan, Seidler, Paul, Rodriguez, Jose, Reyes, Francis E., Sawaya, Michael R., Cascio, Duilio, Weiss, Simon C., Kim, Sun Kyung, Hinck, Cynthia S., Hinck, Andrew P., Calero, Guillermo, Eisenberg, David, and Gonen, Tamir. 2017. "Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED". United States. doi:10.1038/nmeth.4178. https://www.osti.gov/servlets/purl/1463137.
@article{osti_1463137,
title = {Atomic-resolution structures from fragmented protein crystals with the cryoEM method MicroED},
author = {de la Cruz, M. Jason and Hattne, Johan and Shi, Dan and Seidler, Paul and Rodriguez, Jose and Reyes, Francis E. and Sawaya, Michael R. and Cascio, Duilio and Weiss, Simon C. and Kim, Sun Kyung and Hinck, Cynthia S. and Hinck, Andrew P. and Calero, Guillermo and Eisenberg, David and Gonen, Tamir},
abstractNote = {Crystallographic analysis of macromolecules depends on large, well-ordered crystals, which often require significant effort to obtain. Even sizable crystals sometimes suffer from pathologies that render them inappropriate for high-resolution structure determination. Here we show that fragmentation of large, imperfect crystals can provide a simple path for high-resolution structure determination by serial femtosecond crystallography or the cryoEM method MicroED.},
doi = {10.1038/nmeth.4178},
journal = {Nature Methods},
number = 4,
volume = 14,
place = {United States},
year = {2017},
month = {2}
}

Works referenced in this record:

Serial femtosecond crystallography: the first five years
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MOLREP an Automated Program for Molecular Replacement
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SHELXT – Integrated space-group and crystal-structure determination
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Nanoflow electrospinning serial femtosecond crystallography
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Micro-Electron Diffraction data from Tau VQIVYK peptide, source of 5k7n structure
dataset, April 2016
  • Hattne, Johan; Rodriguez, Jose A.; Gonen, Tamir
  • SBGrid Data Bank, Vol. 1
  • DOI: 10.15785/sbgrid/284

Micro-Electron Diffraction data from Lysozyme, source of 5k7o structure
dataset, March 2016
  • Hattne, Johan; Cruz, Jason de la; Gonen, Tamir
  • SBGrid Data Bank, Vol. 1
  • DOI: 10.15785/sbgrid/285

Micro-Electron Diffraction data from Xylanase, source of 5k7p structure
dataset, April 2015
  • Hattne, Johan; Cruz, Jason de la; Gonen, Tamir
  • SBGrid Data Bank, Vol. 1
  • DOI: 10.15785/sbgrid/286

Micro-Electron Diffraction data from Thaumatin, source of 5k7q structure
dataset, March 2015
  • Hattne, Johan; Cruz, Jason de la; Gonen, Tamir
  • SBGrid Data Bank, Vol. 1
  • DOI: 10.15785/sbgrid/287

Micro-Electron Diffraction data from Trypsin, source of 5k7r structure
dataset, March 2016
  • Hattne, Johan; Cruz, Jason de la; Gonen, Tamir
  • SBGrid Data Bank, Vol. 1
  • DOI: 10.15785/sbgrid/288

Micro-Electron Diffraction data from Proteinase K, source of 5k7s structure
dataset, March 2016
  • Hattne, Johan; Cruz, Jason de la; Gonen, Tamir
  • SBGrid Data Bank, Vol. 1
  • DOI: 10.15785/sbgrid/289

Micro-Electron Diffraction data from Thermolysin, source of 5k7t structure
dataset, April 2015
  • Hattne, Johan; Cruz, Jason de la; Gonen, Tamir
  • SBGrid Data Bank, Vol. 1
  • DOI: 10.15785/sbgrid/290