Nature Communications (Dec 2023)
A multi-reservoir extruder for time-resolved serial protein crystallography and compound screening at X-ray free-electron lasers
- Maximilian Wranik,
- Michal W. Kepa,
- Emma V. Beale,
- Daniel James,
- Quentin Bertrand,
- Tobias Weinert,
- Antonia Furrer,
- Hannah Glover,
- Dardan Gashi,
- Melissa Carrillo,
- Yasushi Kondo,
- Robin T. Stipp,
- Georgii Khusainov,
- Karol Nass,
- Dmitry Ozerov,
- Claudio Cirelli,
- Philip J. M. Johnson,
- Florian Dworkowski,
- John H. Beale,
- Scott Stubbs,
- Thierry Zamofing,
- Marco Schneider,
- Kristina Krauskopf,
- Li Gao,
- Oliver Thorn-Seshold,
- Christoph Bostedt,
- Camila Bacellar,
- Michel O. Steinmetz,
- Christopher Milne,
- Jörg Standfuss
Affiliations
- Maximilian Wranik
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Michal W. Kepa
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Emma V. Beale
- Laboratory for Synchrotron Radiation and Femtochemistry, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Daniel James
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Quentin Bertrand
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Tobias Weinert
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Antonia Furrer
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Hannah Glover
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Dardan Gashi
- Laboratory for Synchrotron Radiation and Femtochemistry, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Melissa Carrillo
- Laboratory of Nanoscale Biology, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Yasushi Kondo
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Robin T. Stipp
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Georgii Khusainov
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Karol Nass
- Laboratory for Macromolecules and Bioimaging, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Dmitry Ozerov
- Scientific Computing, Theory and Data Division, Paul Scherrer Institut, Villigen-PSI
- Claudio Cirelli
- Laboratory for Synchrotron Radiation and Femtochemistry, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Philip J. M. Johnson
- Laboratory for Nonlinear Optics, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Florian Dworkowski
- Laboratory for Macromolecules and Bioimaging, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- John H. Beale
- Laboratory for Macromolecules and Bioimaging, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Scott Stubbs
- Large Research Facilities Division, Paul Scherrer Institut, Villigen-PSI
- Thierry Zamofing
- Large Research Facilities Division, Paul Scherrer Institut, Villigen-PSI
- Marco Schneider
- Large Research Facilities Division, Paul Scherrer Institut, Villigen-PSI
- Kristina Krauskopf
- Department of Pharmacy, Ludwig-Maximilians University of Munich
- Li Gao
- Department of Pharmacy, Ludwig-Maximilians University of Munich
- Oliver Thorn-Seshold
- Department of Pharmacy, Ludwig-Maximilians University of Munich
- Christoph Bostedt
- Laboratory for Synchrotron Radiation and Femtochemistry, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Camila Bacellar
- Laboratory for Synchrotron Radiation and Femtochemistry, Photon Science Division, Paul Scherrer Institut, Villigen-PSI
- Michel O. Steinmetz
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- Christopher Milne
- Femtosecond X-ray Experiments Instrument, European XFEL GmbH
- Jörg Standfuss
- Laboratory of Biomolecular Research, Division of Biology and Chemistry, Paul Scherrer Institut, Villigen-PSI
- DOI
- https://doi.org/10.1038/s41467-023-43523-5
- Journal volume & issue
-
Vol. 14,
no. 1
pp. 1 – 12
Abstract
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 supported sample stability. 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 identify 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.
