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Title: Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals

Abstract

Innovative new crystallographic methods are facilitating structural studies from ever smaller crystals of biological macromolecules. In particular, serial X-ray crystallography and microcrystal electron diffraction (MicroED) have emerged as useful methods for obtaining structural information from crystals on the nanometre to micrometre scale. Despite the utility of these methods, their implementation can often be difficult, as they present many challenges that are not encountered in traditional macromolecular crystallography experiments. Here, XFEL serial crystallography experiments and MicroED experiments using batch-grown microcrystals of the enzyme cyclophilin A are described. The results provide a roadmap for researchers hoping to design macromolecular microcrystallography experiments, and they highlight the strengths and weaknesses of the two methods. Specifically, we focus on how the different physical conditions imposed by the sample-preparation and delivery methods required for each type of experiment affect the crystal structure of the enzyme.

Authors:
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Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES). Scientific User Facilities Division; National Science Foundation (NSF); National Institutes of Health (NIH)
OSTI Identifier:
1617957
Alternate Identifier(s):
OSTI ID: 1615313; OSTI ID: 1617040
Grant/Contract Number:  
AC02-05CH11231; AC02-76SF00515; STC-1231306; F32 HL129989; GM123159; GM124149; GM117126; LFR-17-476732
Resource Type:
Journal Article: Published Article
Journal Name:
IUCrJ
Additional Journal Information:
Journal Name: IUCrJ Journal Volume: 7 Journal Issue: 2; Journal ID: ISSN 2052-2525
Publisher:
International Union of Crystallography
Country of Publication:
United Kingdom
Language:
English
Subject:
36 MATERIALS SCIENCE; 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; microcrystals; batch crystallization; serial crystallography; MicroED

Citation Formats

Wolff, Alexander M., Young, Iris D., Sierra, Raymond G., Brewster, Aaron S., Martynowycz, Michael W., Nango, Eriko, Sugahara, Michihiro, Nakane, Takanori, Ito, Kazutaka, Aquila, Andrew, Bhowmick, Asmit, Biel, Justin T., Carbajo, Sergio, Cohen, Aina E., Cortez, Saul, Gonzalez, Ana, Hino, Tomoya, Im, Dohyun, Koralek, Jake D., Kubo, Minoru, Lazarou, Tomas S., Nomura, Takashi, Owada, Shigeki, Samelson, Avi J., Tanaka, Tomoyuki, Tanaka, Rie, Thompson, Erin M., van den Bedem, Henry, Woldeyes, Rahel A., Yumoto, Fumiaki, Zhao, Wei, Tono, Kensuke, Boutet, Sebastien, Iwata, So, Gonen, Tamir, Sauter, Nicholas K., Fraser, James S., and Thompson, Michael C. Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals. United Kingdom: N. p., 2020. Web. doi:10.1107/S205225252000072X.
Wolff, Alexander M., Young, Iris D., Sierra, Raymond G., Brewster, Aaron S., Martynowycz, Michael W., Nango, Eriko, Sugahara, Michihiro, Nakane, Takanori, Ito, Kazutaka, Aquila, Andrew, Bhowmick, Asmit, Biel, Justin T., Carbajo, Sergio, Cohen, Aina E., Cortez, Saul, Gonzalez, Ana, Hino, Tomoya, Im, Dohyun, Koralek, Jake D., Kubo, Minoru, Lazarou, Tomas S., Nomura, Takashi, Owada, Shigeki, Samelson, Avi J., Tanaka, Tomoyuki, Tanaka, Rie, Thompson, Erin M., van den Bedem, Henry, Woldeyes, Rahel A., Yumoto, Fumiaki, Zhao, Wei, Tono, Kensuke, Boutet, Sebastien, Iwata, So, Gonen, Tamir, Sauter, Nicholas K., Fraser, James S., & Thompson, Michael C. Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals. United Kingdom. https://doi.org/10.1107/S205225252000072X
Wolff, Alexander M., Young, Iris D., Sierra, Raymond G., Brewster, Aaron S., Martynowycz, Michael W., Nango, Eriko, Sugahara, Michihiro, Nakane, Takanori, Ito, Kazutaka, Aquila, Andrew, Bhowmick, Asmit, Biel, Justin T., Carbajo, Sergio, Cohen, Aina E., Cortez, Saul, Gonzalez, Ana, Hino, Tomoya, Im, Dohyun, Koralek, Jake D., Kubo, Minoru, Lazarou, Tomas S., Nomura, Takashi, Owada, Shigeki, Samelson, Avi J., Tanaka, Tomoyuki, Tanaka, Rie, Thompson, Erin M., van den Bedem, Henry, Woldeyes, Rahel A., Yumoto, Fumiaki, Zhao, Wei, Tono, Kensuke, Boutet, Sebastien, Iwata, So, Gonen, Tamir, Sauter, Nicholas K., Fraser, James S., and Thompson, Michael C. Wed . "Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals". United Kingdom. https://doi.org/10.1107/S205225252000072X.
@article{osti_1617957,
title = {Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals},
author = {Wolff, Alexander M. and Young, Iris D. and Sierra, Raymond G. and Brewster, Aaron S. and Martynowycz, Michael W. and Nango, Eriko and Sugahara, Michihiro and Nakane, Takanori and Ito, Kazutaka and Aquila, Andrew and Bhowmick, Asmit and Biel, Justin T. and Carbajo, Sergio and Cohen, Aina E. and Cortez, Saul and Gonzalez, Ana and Hino, Tomoya and Im, Dohyun and Koralek, Jake D. and Kubo, Minoru and Lazarou, Tomas S. and Nomura, Takashi and Owada, Shigeki and Samelson, Avi J. and Tanaka, Tomoyuki and Tanaka, Rie and Thompson, Erin M. and van den Bedem, Henry and Woldeyes, Rahel A. and Yumoto, Fumiaki and Zhao, Wei and Tono, Kensuke and Boutet, Sebastien and Iwata, So and Gonen, Tamir and Sauter, Nicholas K. and Fraser, James S. and Thompson, Michael C.},
abstractNote = {Innovative new crystallographic methods are facilitating structural studies from ever smaller crystals of biological macromolecules. In particular, serial X-ray crystallography and microcrystal electron diffraction (MicroED) have emerged as useful methods for obtaining structural information from crystals on the nanometre to micrometre scale. Despite the utility of these methods, their implementation can often be difficult, as they present many challenges that are not encountered in traditional macromolecular crystallography experiments. Here, XFEL serial crystallography experiments and MicroED experiments using batch-grown microcrystals of the enzyme cyclophilin A are described. The results provide a roadmap for researchers hoping to design macromolecular microcrystallography experiments, and they highlight the strengths and weaknesses of the two methods. Specifically, we focus on how the different physical conditions imposed by the sample-preparation and delivery methods required for each type of experiment affect the crystal structure of the enzyme.},
doi = {10.1107/S205225252000072X},
url = {https://www.osti.gov/biblio/1617957}, journal = {IUCrJ},
issn = {2052-2525},
number = 2,
volume = 7,
place = {United Kingdom},
year = {2020},
month = {2}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at https://doi.org/10.1107/S205225252000072X

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