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Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography

Mehrabi, P.; Sung, S.; von Stetten, D.; Prester, A.; Hatton, C. E.; Kleine-Doepke, S.; Berkes, A.; Gore, G.; Leimkohl, J.-P.; Schikora, H.; Kollewe, M.; Rohde, H.; Wilmanns, M.; Tellkamp, F.; Schulz, E. C.

2022-09-21 biophysics
10.1101/2022.09.20.508674 bioRxiv
Show abstract

We introduce the spitrobot, a protein crystal plunger, enabling reaction quenching via cryo-trapping with millisecond time-resolution. Canonical micromesh loops are mounted on an electropneumatic piston, reactions are initiated via the liquid application method (LAMA), and finally intermediate states are cryo-trapped in liquid nitrogen. We demonstrate binding of several ligands in microcrystals of three enzymes, and trapping of reaction intermediates and conformational changes in macroscopic crystals of tryptophan synthase.

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