8G31

Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate

これはPDB形式変換不可エントリーです。

8G31 の概要

• エントリーDOI: 10.2210/pdb8g31/pdb
• EMDBエントリー: 29681 29687 29688 29689 29724 29844
• 分子名称: 50S ribosomal protein L32, 50S ribosomal protein L3, 50S ribosomal protein L4, ... (52 entities in total)
• 機能のキーワード: recycling, time-resolved cryo-em, 70s, hflx, ribosome
• 由来する生物種: Escherichia coli; 詳細
• タンパク質・核酸の鎖数: 52
• 化学式量合計: 2140230.47

構造登録者

Bhattacharjee, S.,Brown, P.Z.,Frank, J. (登録日: 2023-02-06, 公開日: 2023-12-06, 最終更新日: 2024-02-14)

主引用文献

Bhattacharjee, S.,Feng, X.,Maji, S.,Dadhwal, P.,Zhang, Z.,Brown, Z.P.,Frank, J.
Time resolution in cryo-EM using a PDMS-based microfluidic chip assembly and its application to the study of HflX-mediated ribosome recycling.
Cell, 187:782-796.e23, 2024

PubMed Abstract: The rapid kinetics of biological processes and associated short-lived conformational changes pose a significant challenge in attempts to structurally visualize biomolecules during a reaction in real time. Conventionally, on-pathway intermediates have been trapped using chemical modifications or reduced temperature, giving limited insights. Here, we introduce a time-resolved cryo-EM method using a reusable PDMS-based microfluidic chip assembly with high reactant mixing efficiency. Coating of PDMS walls with SiO virtually eliminates non-specific sample adsorption and ensures maintenance of the stoichiometry of the reaction, rendering it highly reproducible. In an operating range from 10 to 1,000 ms, the device allows us to follow in vitro reactions of biological molecules at resolution levels in the range of 3 Å. By employing this method, we show the mechanism of progressive HflX-mediated splitting of the 70S E. coli ribosome in the presence of the GTP via capture of three high-resolution reaction intermediates within 140 ms.

PubMed: 38244547
DOI: 10.1016/j.cell.2023.12.027

実験手法

ELECTRON MICROSCOPY (3.2 Å)