8BK7

Cryo-EM structure of beta-galactosidase at 3.3 A resolution plunged 5 ms after mixing with apoferritin

8BK7 の概要

• エントリーDOI: 10.2210/pdb8bk7/pdb
• EMDBエントリー: 16091
• 分子名称: Beta-galactosidase (1 entity in total)
• 機能のキーワード: glycosyl hydrolase, hydrolase
• 由来する生物種: Escherichia coli K-12
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 469953.50

構造登録者

Torino, S.,Dhurandhar, M.,Efremov, R. (登録日: 2022-11-08, 公開日: 2023-08-09, 最終更新日: 2023-09-13)

主引用文献

Torino, S.,Dhurandhar, M.,Stroobants, A.,Claessens, R.,Efremov, R.G.
Time-resolved cryo-EM using a combination of droplet microfluidics with on-demand jetting.
Nat.Methods, 20:1400-1408, 2023

PubMed Abstract: Single-particle cryogenic electron microscopy (cryo-EM) allows reconstruction of high-resolution structures of proteins in different conformations. Protein function often involves transient functional conformations, which can be resolved using time-resolved cryo-EM (trEM). In trEM, reactions are arrested after a defined delay time by rapid vitrification of protein solution on the EM grid. Despite the increasing interest in trEM among the cryo-EM community, making trEM samples with a time resolution below 100 ms remains challenging. Here we report the design and the realization of a time-resolved cryo-plunger that combines a droplet-based microfluidic mixer with a laser-induced generator of microjets that allows rapid reaction initiation and plunge-freezing of cryo-EM grids. Using this approach, a time resolution of 5 ms was achieved and the protein density map was reconstructed to a resolution of 2.1 Å. trEM experiments on GroEL:GroES chaperonin complex resolved the kinetics of the complex formation and visualized putative short-lived conformations of GroEL-ATP complex.

PubMed: 37592181
DOI: 10.1038/s41592-023-01967-z

実験手法

ELECTRON MICROSCOPY (3.3 Å)