9R4H

Spitrobot-2 advances time-resolvedcryo-trapping crystallography to under 25 ms: T4 Lysozyme, mutant L99A bound with indole (10 s soaking)

Summary for 9R4H

• Entry DOI: 10.2210/pdb9r4h/pdb
• Descriptor: Endolysin, INDOLE, SODIUM ION, ... (4 entities in total)
• Functional Keywords: t4 lysozyme, l99a, hydrolase
• Biological source: Escherichia phage T4
• Total number of polymer chains: 1
• Total formula weight: 18646.20

Authors

Spiliopoulou, M.,Hatton, C.E.,Mehrabi, P.,Schulz, E.C. (deposition date: 2025-05-07, release date: 2025-12-03)

Primary citation

Spiliopoulou, M.,Hatton, C.E.,Kollewe, M.,Leimkohl, J.P.,Schikora, H.,Tellkamp, F.,Mehrabi, P.,Schulz, E.C.
Spitrobot-2 advances time-resolved cryo-trapping crystallography to under 25 ms.
Commun Chem, 8:363-363, 2025

PubMed 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.

PubMed: 41266557
DOI: 10.1038/s42004-025-01784-9

Experimental method

X-RAY DIFFRACTION (1.49 Å)