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

This paper introduces the spitrobot, a protein crystal plunger enabling reaction quenching via cryo-trapping with millisecond time resolution. Protein crystals are mounted on micromeshes, reactions initiated via liquid application, and plunging occurs after a set delay. High-magnification images are automatically recorded, and the SPINE-standard holder ensures high-throughput compatibility. The authors demonstrate its use in studying glucose and 2,3-butanediol binding in xylose isomerase, avibactam and ampicillin binding in CTX-M-14 β-lactamase, and trapping reaction intermediates in tryptophan synthase, showcasing its potential for studying catalytic events.

Pedram Mehrabi, Sihyun Sung, David von Stetten, Andreas Prester, Caitlin E. Hatton, Stephan Kleine-Döpke, Alexander Berkes, Gargi Gore, Jan-Philipp Leimkohl, Hendrik Schikora, Martin Kollewe, Holger Rohde, Matthias Wilmanns, Friedjof Tellkamp, Eike C. Schulz