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

IUCrJ
Alexander M WolffMichael C Thompson

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.

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Citations

Jul 18, 2020·F1000Research·Michael C ThompsonJose A Rodriguez
Aug 2, 2020·Communications Biology·Max T B ClabbersHongyi Xu
Dec 10, 2020·Proceedings of the National Academy of Sciences of the United States of America·Michael W MartynowyczTamir Gonen
Jan 9, 2021·Quarterly Reviews of Biophysics·Marcus Fischer
Oct 3, 2020·Structure·Michael W Martynowycz, Tamir Gonen
Mar 2, 2021·Acta Crystallographica. Section D, Structural Biology·Max T B Clabbers, Hongyi Xu
May 5, 2021·Acta Crystallographica. Section A, Foundations and Advances·R BeanlandS Kamba
Jul 3, 2021·Toxins·Guillaume TetreauJacques-Philippe Colletier
Nov 3, 2021·Acta Crystallographica. Section F, Structural Biology Communications·Matthew W Bowler

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Methods Mentioned

BETA
X-ray
electron diffraction
electron cryomicroscopy
light microscopy
light-scattering
electron scattering

Software Mentioned

Fiji
Coot
cctbx
xfel
DirAx
Phaser
PyMOL
CrystFEL
merge
phenix

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