Comparing serial X-ray crystallography and microcrystal electron diffraction (MicroED) as methods for routine structure determination from small macromolecular crystals
- Creators
- Wolff, Alexander M.
- Zhao, Wei
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.
Additional Information
© 2020 International Union of Crystallography. This is an open-access article distributed under the terms of the Creative Commons Attribution (CC-BY) Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited. Received 13 September 2019; Accepted 21 January 2020. We thank J. Rodriguez and D. Hekstra for helpful insight. FIB milling was performed in the Beckman Institute Resource Center at the California Institute of Technology. Portions of this research were carried out under proposals LQ79, P074, and LO19 at the Linac Coherent Light Source (LCLS) at the SLAC National Accelerator Laboratory, supported by the DOE Office of Science, OBES under contract DE-AC02-76SF00515. The HERA system for experiments at MFX was developed by Bruce Doak and is funded by the Max-Planck Institute for Medical Research. Portions of this research were performed on beamline 3 at SACLA with the approval of the Japan Synchrotron Radiation Research Institute (JASRI; proposal No. 2017B8055). We thank the staff at SACLA for their assistance. Data processing was performed in part at the National Energy Research Scientific Computing Center, supported by the DOE Office of Science, Contract No. DEAC02-05CH11231. MCT is supported by NSF STC-1231306, a Ruth L. Kirschstein National Research Service Award (F32 HL129989) and the UCSF Program in Breakthrough Biomedical Sciences. JSF is supported by a Packard Fellowship from the David and Lucile Packard Foundation, NIH GM123159, NIH GM124149, UC Office of the President Laboratory Fees Research Program LFR-17-476732 and NSF STC-1231306. NKS is supported by NIH GM117126. SI is supported by the Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research; BINDS) from the Japan Agency for Medical Research and Development (AMED). RAW was supported by the NSF Graduate Research Fellowship.Attached Files
Published - mf5038.pdf
Submitted - 767061.full.pdf
Supplemental Material - mf5038sup1.pdf
Files
Additional details
- PMCID
- PMC7055375
- Eprint ID
- 98636
- Resolver ID
- CaltechAUTHORS:20190913-092049265
- DBI-1231306
- NSF
- F32 HL129989
- NIH Postdoctoral Fellowship
- University of California, San Francisco
- David and Lucile Packard Foundation
- GM123159
- NIH
- GM124149
- NIH
- LFR-17-476732
- University of California Lab Fees Research Program
- STC-1231306
- NSF
- GM117126
- NIH
- Japan Agency for Medical Research and Development
- NSF Graduate Research Fellowship
- DE-AC02-76SF00515
- Department of Energy (DOE)
- Max-Planck Institute for Medical Research
- 2017B8055
- Japan Synchrotron Radiation Research Institute
- DEAC02-05CH11231
- Department of Energy (DOE)
- Created
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2019-09-13Created from EPrint's datestamp field
- Updated
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2023-09-08Created from EPrint's last_modified field