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A multi-reservoir extruder for time-resolved serial protein crystallography and compound screening at X-ray free-electron lasers
Open accessPreprint — published as 10.1038/s41467-023-43523-5 · Paper page
Abstract
Serial crystallography at X-ray free-electron lasers (XFELs) permits the determination of radiation-damage free static as well as time-resolved protein structures at room temperature. Efficient sample delivery is a key factor for such experiments. Here, we describe a multi-reservoir, high viscosity extruder as a step towards automation of sample delivery at XFELs. Compared to a standard single extruder, sample exchange time was halved and the workload of users was greatly reduced. In-built temperature control of samples facilitated optimal extrusion and offered a path forward for the observation of temperature-dependent structural changes. After commissioning the device with lysozyme crystals, we collected time-resolved data using crystals of a membrane-bound, light-driven sodium pump. Static data were also collected from the soluble protein tubulin that was soaked with a series of small molecule drugs. Using these data, we identified low occupancy (as little as 30%) ligands using a minimal amount of data from a serial crystallography experiment, a result that could be exploited for structure-based drug design.
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Cites
Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764
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Metadata
- Authors
- Wranik, M., Kepa, M., Beale, E., James, D., Bertrand, Q., Weinert, T., Furrer, A., Glover, H., Gashi, D., Carrillo, M., Kondo, Y., Stipp, R., Khusainov, G., Nass, K., Ozerov, D., Cirelli, C., Johnson, P., Dworkowski, F., Beale, J., Stubbs, S., Zamofing, T., Schneider, M., Krauskopf, K., Gao, L., Thorn-Seshold, O., Bostedt, C., Bacellar, C., Steinmetz, M., Milne, C., & Standfuss, J.
- Affiliations
- Paul Scherrer Institute (CH)
- Ludwig-Maximilians-Universität München (DE)
- Ludwig-Maximilians University of Munich
- University of Basel (CH)
- European X-Ray Free-Electron Laser (DE)
- Corresponding author
- Wranik, Maximilian
- Kepa, Michal
- ORCID
- Wranik, Maximilian
- Kepa, Michal
- Beale, Emma
- James, Daniel
- Bertrand, Quentin
- Weinert, Tobias
- Furrer, Antonia
- Glover, Hannah
- Carrillo, Melissa
- Kondo, Yasushi
- Stipp, Robin
- Khusainov, Georgii
- Nass, Karol
- Ozerov, Dmitry
- Cirelli, Claudio
- Johnson, Philip
- Dworkowski, Florian
- Beale, John
- Schneider, Marco
- Krauskopf, Kristina
- Gao, Li
- Thorn-Seshold, Oliver
- Bostedt, Christoph
- Bacellar, Camila
- Steinmetz, Michel
- Milne, Christopher
- Standfuss, Jörg
- Venue
- Nature Communications
- Published
- 2023-03-09
- Volume
- 14
- Issue
- 1
- Pages
- 7956-7956
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC10693631
- OpenAlex
- W4389264817
- Cited by
- 14
- References
- 1
- Citations by year
- 2026: 5 · 2025: 6 · 2024: 3
- FWCI
- 1.03
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- Research Square (acceptedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DORA PSI (Paul Scherrer Institute) (submittedVersion, cc-by)
- Infoscience (Ecole Polytechnique Fédérale de Lausanne) (submittedVersion, cc-by-nc-nd)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- European XFEL (submittedVersion, cc-by)
- Funders
- National Science Foundation
- Deutsche Forschungsgemeinschaft (TH2231/1-1, 400324123)
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (207462, 179351, 192566, 310030)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Enzyme Structure and Function
- Physical Sciences › Physics and Astronomy › Radiation › Advanced X-ray Imaging Techniques
- Physical Sciences › Chemistry › Inorganic Chemistry › Crystal structures of chemical compounds
- Keywords
- time-resolved serial crystallography, X-ray free-electron laser, protein crystallography, sample temperature control, lysozyme crystals, membrane proteins, tubulin, ligand occupancy, structure-based drug design, compound screening
- MeSH terms
- Crystallography, Electrons, Lasers, Proteins, Synchrotrons, Crystallography, X-Ray
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