8B05

TRYPTOPHAN SYNTHASE - Cryo-trapping by the spitrobot crystal plunger after 20 sec

8B05 の概要

• エントリーDOI: 10.2210/pdb8b05/pdb
• 分子名称: Tryptophan synthase alpha chain, Tryptophan synthase beta chain, CESIUM ION, ... (7 entities in total)
• 機能のキーワード: trytophan synthase, time-resolved crystallography, lyase
• 由来する生物種: Salmonella enterica subsp. enterica serovar Typhimurium; 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 72310.32

構造登録者

Mehrabi, P.,Sung, S.,von Stetten, D.,Prester, A.,Hatton, C.E.,Kleine-Dopke, S.,Berkes, A.,Gore, G.,Leimkohl, J.P.,Schikora, H.,Kollewe, M.,Rohde, H.,Wilmanns, M.,Tellkamp, F.,Schulz, E.C. (登録日: 2022-09-07, 公開日: 2023-05-03, 最終更新日: 2024-02-07)

主引用文献

Mehrabi, P.,Sung, S.,von Stetten, D.,Prester, A.,Hatton, C.E.,Kleine-Dopke, S.,Berkes, A.,Gore, G.,Leimkohl, J.P.,Schikora, H.,Kollewe, M.,Rohde, H.,Wilmanns, M.,Tellkamp, F.,Schulz, E.C.
Millisecond cryo-trapping by the spitrobot crystal plunger simplifies time-resolved crystallography.
Nat Commun, 14:2365-2365, 2023

PubMed Abstract: 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.

PubMed: 37185266
DOI: 10.1038/s41467-023-37834-w

実験手法

X-RAY DIFFRACTION (2.1 Å)