Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography
Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography
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.
Gore Gargi、Leimkohl Jan-Philipp、Kollewe Martin、Tellkamp Friedjof、Schulz Eike C.、Sung Sihyun、von Stetten David、Prester Andreas、Mehrabi Pedram、Kleine-Doepke Stephan、Hatton Caitlin E.、Berkes Alexander、Wilmanns Matthias、Rohde Holger、Schikora Hendrik
晶体学生物科学研究方法、生物科学研究技术生物化学
Gore Gargi,Leimkohl Jan-Philipp,Kollewe Martin,Tellkamp Friedjof,Schulz Eike C.,Sung Sihyun,von Stetten David,Prester Andreas,Mehrabi Pedram,Kleine-Doepke Stephan,Hatton Caitlin E.,Berkes Alexander,Wilmanns Matthias,Rohde Holger,Schikora Hendrik.Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography[EB/OL].(2025-03-28)[2025-06-29].https://www.biorxiv.org/content/10.1101/2022.09.20.508674.点此复制
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