Serial protein crystallography in an electron microscope

Serial X-ray crystallography at free-electron lasers allows to solve biomolecular structures from sub-micron-sized crystals. However, beam time at these facilities is scarce, and involved sample delivery techniques are required. On the other hand, rotation electron diffraction (MicroED) has shown great potential as an alternative means for protein nanocrystallography. Here, we present a method for serial electron diffraction of protein nanocrystals combining the benefits of both approaches. In a scanning transmission electron microscope, crystals randomly dispersed on a sample grid are automatically mapped, and a diffraction pattern at fixed orientation is recorded from each at a high acquisition rate. Dose fractionation ensures minimal radiation damage effects. We demonstrate the method by solving the structure of granulovirus occlusion bodies and lysozyme to resolutions of 1.55 Å and 1.80 Å, respectively. Our method promises to provide rapid structure determination for many classes of materials with minimal sample consumption, using readily available instrumentation.

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004: Informatik

More Funding Information

Sprovided by the Max Planck Society, Deutsches Elektronen-Synchrotron (DESY), the excellence cluster “The Hamburg Center for Ultrafast Imaging” of the Deutsche Forschungsgemeinschaft (EXC 1074 project ID 194651731), the European Research Council project “Attosecond X-ray Science: Imaging and Spectroscopy” (Award/Contract Number ERC-2013-SyG 609920), and the Joachim Herz Foundation (Biomedical Physics of Infection). P.H. acknowledges support by the Natural Sciences and Engineering Research Council of Canada. P.M. was supported by the Alexander von Humboldt-Stiftung for postdoctoral researchers.

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https://creativecommons.org/licenses/by/4.0/

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s41467-020-14793-0.pdf

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1.67 MB

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Serial protein crystallography in an electron microscope