TY  - JOUR
AU  - Mehrabi, Pedram
AU  - Sung, Sihyun
AU  - Stetten, David von
AU  - Prester, Andreas
AU  - Hatton, Caitlin E.
AU  - Kleine-Döpke, Stephan
AU  - Berkes, Alexander
AU  - Gore, Gargi
AU  - Leimkohl, Jan-Philipp
AU  - Schikora, Hendrik
AU  - Kollewe, Martin
AU  - Rohde, Holger
AU  - Wilmanns, Matthias
AU  - Tellkamp, Friedjof
AU  - Schulz, Eike
TI  - Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography
JO  - Nature Communications
VL  - 14
IS  - 1
SN  - 2041-1723
CY  - [London]
PB  - Nature Publishing Group UK
M1  - PUBDB-2023-02014
SP  - 2365
PY  - 2023
AB  - 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.
LB  - PUB:(DE-HGF)16
C6  - 37185266
UR  - <Go to ISI:>//WOS:001003181400017
DO  - DOI:10.1038/s41467-023-37834-w
UR  - https://bib-pubdb1.desy.de/record/582469
ER  -