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Kilohertz serial crystallography with the JUNGFRAU detector at a fourth-generation synchrotron source

Leonarski, Filip ; Nan, Jie LU ; Matej, Zdenek LU orcid ; Bertrand, Quentin ; Furrer, Antonia ; Gorgisyan, Ishkhan LU ; Bjelčić, Monika LU ; Kepa, Michal ; Glover, Hannah and Hinger, Viktoria , et al. (2023) In IUCrJ 10(Pt 6). p.729-737
Abstract

Serial and time-resolved macromolecular crystallography are on the rise. However, beam time at X-ray free-electron lasers is limited and most thirdgeneration synchrotron-based macromolecular crystallography beamlines do not offer the necessary infrastructure yet. Here, a new setup is demonstrated, based on the JUNGFRAU detector and Jungfraujoch data-acquisition system, that enables collection of kilohertz serial crystallography data at fourthgeneration synchrotrons. More importantly, it is shown that this setup is capable of collecting multiple-time-point time-resolved protein dynamics at kilohertz rates, allowing the probing of microsecond to second dynamics at synchrotrons in a fraction of the time needed previously. A high-quality... (More)

Serial and time-resolved macromolecular crystallography are on the rise. However, beam time at X-ray free-electron lasers is limited and most thirdgeneration synchrotron-based macromolecular crystallography beamlines do not offer the necessary infrastructure yet. Here, a new setup is demonstrated, based on the JUNGFRAU detector and Jungfraujoch data-acquisition system, that enables collection of kilohertz serial crystallography data at fourthgeneration synchrotrons. More importantly, it is shown that this setup is capable of collecting multiple-time-point time-resolved protein dynamics at kilohertz rates, allowing the probing of microsecond to second dynamics at synchrotrons in a fraction of the time needed previously. A high-quality complete X-ray dataset was obtained within 1 min from lysozyme microcrystals, and the dynamics of the light-driven sodium-pump membrane protein KR2 with a time resolution of 1 ms could be demonstrated. To make the setup more accessible for researchers, downstream data handling and analysis will be automated to allow on-the-fly spot finding and indexing, as well as data processing.

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@article{846c1597-ad41-437a-bb2c-5d19e79ebbd7,
  abstract     = {{<p>Serial and time-resolved macromolecular crystallography are on the rise. However, beam time at X-ray free-electron lasers is limited and most thirdgeneration synchrotron-based macromolecular crystallography beamlines do not offer the necessary infrastructure yet. Here, a new setup is demonstrated, based on the JUNGFRAU detector and Jungfraujoch data-acquisition system, that enables collection of kilohertz serial crystallography data at fourthgeneration synchrotrons. More importantly, it is shown that this setup is capable of collecting multiple-time-point time-resolved protein dynamics at kilohertz rates, allowing the probing of microsecond to second dynamics at synchrotrons in a fraction of the time needed previously. A high-quality complete X-ray dataset was obtained within 1 min from lysozyme microcrystals, and the dynamics of the light-driven sodium-pump membrane protein KR2 with a time resolution of 1 ms could be demonstrated. To make the setup more accessible for researchers, downstream data handling and analysis will be automated to allow on-the-fly spot finding and indexing, as well as data processing.</p>}},
  author       = {{Leonarski, Filip and Nan, Jie and Matej, Zdenek and Bertrand, Quentin and Furrer, Antonia and Gorgisyan, Ishkhan and Bjelčić, Monika and Kepa, Michal and Glover, Hannah and Hinger, Viktoria and Eriksson, Thomas and Cehovin, Aleksander and Eguiraun, Mikel and Gasparotto, Piero and Mozzanica, Aldo and Weinert, Tobias and Gonzalez, Ana and Standfuss, Jörg and Wang, Meitian and Ursby, Thomas and Dworkowski, Florian}},
  issn         = {{2052-2525}},
  keywords     = {{data networks; fourth-generation synchrotrons; JUNGFRAU detector; macromolecular crystallography; protein structure; time-resolved serial crystallography; X-ray detectors}},
  language     = {{eng}},
  month        = {{10}},
  number       = {{Pt 6}},
  pages        = {{729--737}},
  publisher    = {{International Union of Crystallography}},
  series       = {{IUCrJ}},
  title        = {{Kilohertz serial crystallography with the JUNGFRAU detector at a fourth-generation synchrotron source}},
  url          = {{http://dx.doi.org/10.1107/S2052252523008618}},
  doi          = {{10.1107/S2052252523008618}},
  volume       = {{10}},
  year         = {{2023}},
}