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.

doi:10.1101/2022.09.20.508674

bioRxiv · 10.1101/2022.09.20.508674

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.

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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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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. Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography. https://doi.org/10.1101/2022.09.20.508674

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