Journal Article

PUBDB-2025-05617

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Spitrobot-2 advances time-resolved cryo-trapping crystallography to under 25 ms

Spiliopoulou, M.Extern*EMBL* ; Hatton, C.Extern* ; Kollewe, M. ; Leimkohl, J.-P. ; Schikora, H. ; Tellkamp, F. (Corresponding author)CFEL*MPG* ; Mehrabi, P. (Corresponding author)Extern*MPG* ; Schulz, E. (Corresponding author)Extern*

2025
Macmillan Publishers Limited, part of Springer Nature [London]

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Please use a persistent id in citations: doi:10.1038/s42004-025-01784-9  doi:10.3204/PUBDB-2025-05617

Abstract: We previously introduced the spitrobot, a protein crystal plunging system that enables reaction quenching via cryo-trapping with a time resolution in the millisecond range. Here we present the next generation, spitrobot-2, as an integrated benchtop device. User-friendliness has been improved by semi-automatic sample exchange. Moreover, a fully automated shutter shields the liquid nitrogen from the humidified environment, improving sample integrity. Most importantly, the cryo-trapping delay time has been reduced to 23 ms, making spitrobot-2 twice as fast as the previous generation. This further expands the number of target systems that can be addressed by cryo-trapping time-resolved crystallography. Using 12 crystal structures of three independent model systems, we demonstrate successful cryo-trapping via observation of conformational changes and ligand binding within 25 ms. These improvements increase the convenient access to cryo-trapping, time-resolved X-ray crystallography empowering the MX community with efficient tools to advance research in structural biology.

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Contributing Institute(s):

1. EMBL-User (EMBL-User)

Research Program(s):

1. 6G3 - PETRA III (DESY) (POF4-6G3) (POF4-6G3)
2. DFG project G:(GEPRIS)451079909 - Untersuchung allosterischer Mechanismen durch zeitaufgelöste serielle Synchrotronkristallographie (451079909) (451079909)
3. DFG project G:(GEPRIS)458246365 - Zeitaufgelöste Strukturanalysde der extended spectrum Beta-Lactamase CTX-M-14 (458246365) (458246365)
4. DynaPLIX - Dynamics of Protein–Ligand Interactions (101071843) (101071843)

Experiment(s):

1. PETRA Beamline P14 (PETRA III)

Appears in the scientific report 2025

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