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

Serial and time-resolved macromolecular crystallography are on the rise. However, beam time at X-ray free-electron lasers is limited and most third-generation synchrotron-based macromolecular crystallography beamlines do not offer the necessary infrastructure yet. Here, a new setup is demonstrated,...

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Detalles Bibliográficos
Autores principales: Leonarski, Filip, Nan, Jie, Matej, Zdenek, Bertrand, Quentin, Furrer, Antonia, Gorgisyan, Ishkhan, Bjelčić, Monika, Kepa, Michal, Glover, Hannah, Hinger, Viktoria, Eriksson, Thomas, Cehovin, Aleksander, Eguiraun, Mikel, Gasparotto, Piero, Mozzanica, Aldo, Weinert, Tobias, Gonzalez, Ana, Standfuss, Jörg, Wang, Meitian, Ursby, Thomas, Dworkowski, Florian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Union of Crystallography 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10619449/
https://www.ncbi.nlm.nih.gov/pubmed/37830774
http://dx.doi.org/10.1107/S2052252523008618
Descripción
Sumario:Serial and time-resolved macromolecular crystallography are on the rise. However, beam time at X-ray free-electron lasers is limited and most third-generation 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 fourth-generation 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.