EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Time resolution in cryo-EM using a PDMS-based microfluidic chip assembly and its application to the study of HflX-mediated ribosome recycling.
ジャーナル・号・ページ	Cell, Vol. 187, Issue 3, Page 782-796.e23, Year 2024
掲載日	2024年2月1日
著者	Sayan Bhattacharjee / Xiangsong Feng / Suvrajit Maji / Prikshat Dadhwal / Zhening Zhang / Zuben P Brown / Joachim Frank /
PubMed 要旨	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 ...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.
リンク	Cell / PubMed:38244547 / PubMed Central
手法	EM (単粒子)
解像度	3.0 - 4.1 Å
構造データ	29681 8g2u EMDB-29681, PDB-8g2u: Time-resolved cryo-EM study of the 70S recycling by the HflX:control-apo-70S at 900ms 手法: EM (単粒子) / 解像度: 3.0 Å 29687 8g31 EMDB-29687, PDB-8g31: Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate 手法: EM (単粒子) / 解像度: 3.2 Å 29688 8g34 EMDB-29688, PDB-8g34: Time-resolved cryo-EM study of the 70S recycling by the HflX:1st intermediate 手法: EM (単粒子) / 解像度: 3.2 Å 29689 8g38 EMDB-29689, PDB-8g38: Time-resolved cryo-EM study of the 70S recycling by the HflX:3rd Intermediate 手法: EM (単粒子) / 解像度: 3.2 Å EMDB-29723: Time-resolved cryo-EM study of the 70S recycling by the HflX:3rd Intermediate, 30S focused map 手法: EM (単粒子) / 解像度: 4.1 Å EMDB-29724: Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate consensus, 30S focused 手法: EM (単粒子) / 解像度: 3.4 Å EMDB-29833: Time-resolved cryo-EM study of the 70S recycling by the HflX:1st intermediate, 30S focused 手法: EM (単粒子) / 解像度: 3.2 Å EMDB-29834: Time-resolved cryo-EM study of the 70S recycling by the HflX:1st intermediate, 50S focused from 30S subtracted 手法: EM (単粒子) / 解像度: 3.6 Å EMDB-29842: Time-resolved cryo-EM study of the 70S recycling by the HflX:3rd Intermediate, 50S focused and 30S subtracted 手法: EM (単粒子) / 解像度: 3.0 Å EMDB-29844: Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate, 50S focused and 30S subtracted 手法: EM (単粒子) / 解像度: 3.14 Å
化合物	ChemComp-GTP: GUANOSINE-5'-TRIPHOSPHATE / GTP / GTP, エネルギー貯蔵分子YM ChemComp-HOH*: WATER
由来	escherichia coli (大腸菌)
キーワード	RIBOSOME / Recycling / Time-resolved Cryo-EM / 70S / HflX