An automated platform for in situ serial crystallography at room temperature.

IUCrJ
Zhong RenXiaojing Yang

Abstract

Direct observation of functional motions in protein structures is highly desirable for understanding how these nanomachineries of life operate at the molecular level. Because cryogenic temperatures are non-physiological and may prohibit or even alter protein structural dynamics, it is necessary to develop robust X-ray diffraction methods that enable routine data collection at room temperature. We recently reported a crystal-on-crystal device to facilitate in situ diffraction of protein crystals at room temperature devoid of any sample manipulation. Here an automated serial crystallography platform based on this crystal-on-crystal technology is presented. A hardware and software prototype has been implemented, and protocols have been established that allow users to image, recognize and rank hundreds to thousands of protein crystals grown on a chip in optical scanning mode prior to serial introduction of these crystals to an X-ray beam in a programmable and high-throughput manner. This platform has been tested extensively using fragile protein crystals. We demonstrate that with affordable sample consumption, this in situ serial crystallography technology could give rise to room-temperature protein structures of higher resolution ...Continue Reading

References

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Jun 1, 2017·Proceedings of the National Academy of Sciences of the United States of America·Sepalika BandaraXiaojing Yang
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Sep 19, 2019·Proceedings of the National Academy of Sciences of the United States of America·Heewhan ShinXiaojing Yang

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Citations

Mar 18, 2021·Proceedings of the National Academy of Sciences of the United States of America·Sepalika BandaraXiaojing Yang

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Methods Mentioned

BETA
protein crystallography
X-ray
chips
chip
Bragg diffraction
dynamic crystallography
size-exclusion chromatography

Software Mentioned

PHENIX
Precognition
inSituX
Coot
Open Source Computer Vision Library OpenCV
/ Epinorm

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