Polyacrylamide injection matrix for serial femtosecond crystallography

Scientific Reports
Jaehyun ParkKi Hyun Nam

Abstract

Serial femtosecond crystallography (SFX) provides opportunities to observe the dynamics of macromolecules without causing radiation damage at room temperature. Although SFX provides a biologically more reliable crystal structure than provided by the existing synchrotron sources, there are limitations due to the consumption of many crystal samples. A viscous medium as a carrier matrix reduces the flow rate of the crystal sample from the injector, thereby dramatically reducing sample consumption. However, the currently available media cannot be applied to specific crystal samples owing to reactions between the viscous medium and crystal sample. The discovery and characterisation of a new delivery medium for SFX can further expand its use. Herein, we report the preparation of a polyacrylamide (PAM) injection matrix to determine the crystal structure with an X-ray free-electron laser. We obtained 11,936 and 22,213 indexed images using 0.5 mg lysozyme and 1.0 mg thermolysin, respectively. We determined the crystal structures of lysozyme and thermolysin delivered in PAM at 1.7 Å and 1.8 Å resolutions. The maximum background scattering from PAM was lower than monoolein, a commonly used viscous medium. Our results show that PAM can be ...Continue Reading

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Citations

Oct 17, 2019·The FEBS Journal·Feng-Zhu ZhaoDa-Chuan Yin
Sep 7, 2019·Journal of Synchrotron Radiation·Suk Youl Park, Ki Hyun Nam
May 8, 2019·Scientific Reports·Donghyeon LeeKi Hyun Nam
Jun 30, 2019·Analytical and Bioanalytical Chemistry·Austin EchelmeierAlexandra Ros
Aug 23, 2020·International Journal of Molecular Sciences·Ki Hyun Nam
Apr 14, 2020·Journal of Applied Crystallography·Donghyeon LeeJaehyun Park
Mar 7, 2019·International Journal of Molecular Sciences·Ki Hyun Nam
Jan 11, 2020·Scientific Reports·Ki Hyun Nam
Jan 30, 2020·Scientific Reports·Michihiro SugaharaTetsuya Ishikawa
May 14, 2020·International Journal of Molecular Sciences·Ki Hyun Nam
Jan 15, 2022·Scientific Reports·Ki Hyun Nam

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Methods Mentioned

BETA
X-ray
electrophoresis

Software Mentioned

MolProbity
CrystFEL
PHENIX
Coot

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