%0 Journal Article
%A Mehrabi, Pedram
%A Sung, Sihyun
%A Stetten, David von
%A Prester, Andreas
%A Hatton, Caitlin E.
%A Kleine-Döpke, Stephan
%A Berkes, Alexander
%A Gore, Gargi
%A Leimkohl, Jan-Philipp
%A Schikora, Hendrik
%A Kollewe, Martin
%A Rohde, Holger
%A Wilmanns, Matthias
%A Tellkamp, Friedjof
%A Schulz, Eike
%T Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography
%J Nature Communications
%V 14
%N 1
%@ 2041-1723
%C [London]
%I Nature Publishing Group UK
%M PUBDB-2023-02014
%P 2365
%D 2023
%X We introduce the spitrobot, a protein crystal plunger, enabling reaction quenching via cryo-trapping with a time-resolution in the millisecond range. Protein crystals are mounted on canonical micromeshes on an electropneumatic piston, where the crystals are kept in a humidity and temperature-controlled environment, then reactions are initiated via the liquid application method (LAMA) and plunging into liquid nitrogen is initiated after an electronically set delay time to cryo-trap intermediate states. High-magnification images are automatically recorded before and after droplet deposition, prior to plunging. The SPINE-standard sample holder is directly plunged into a storage puck, enabling compatibility with high-throughput infrastructure. Here we demonstrate binding of glucose and 2,3-butanediol in microcrystals of xylose isomerase, and of avibactam and ampicillin in microcrystals of the extended spectrum beta-lactamase CTX-M-14. We also trap reaction intermediates and conformational changes in macroscopic crystals of tryptophan synthase to demonstrate that the spitrobot enables insight into catalytic events.
%F PUB:(DE-HGF)16
%9 Journal Article
%$ 37185266
%U <Go to ISI:>//WOS:001003181400017
%R 10.1038/s41467-023-37834-w
%U https://bib-pubdb1.desy.de/record/582469