Database of articles cited by EMDB/PDB/SASBDB data

Keywords
Structure methods	All X-ray diffraction NMR electron microscopy small angle scattering neutron diffraction fiber diffraction powder diffraction infrared spectroscopy EPR fluorescence transfer theoretical model Hybrid
Author
Journal
IF	>30 >20 >10 >5 all

Title	Time resolution in cryo-EM using a PDMS-based microfluidic chip assembly and its application to the study of HflX-mediated ribosome recycling.
Journal, issue, pages	Cell, Vol. 187, Issue 3, Page 782-796.e23, Year 2024
Publish date	Feb 1, 2024
Authors	Sayan Bhattacharjee / Xiangsong Feng / Suvrajit Maji / Prikshat Dadhwal / Zhening Zhang / Zuben P Brown / Joachim Frank /
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 ...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.
External links	Cell / PubMed:38244547 / PubMed Central
Methods	EM (single particle)
Resolution	3.0 - 4.1 Å
Structure data	29681 8g2u EMDB-29681, PDB-8g2u: Time-resolved cryo-EM study of the 70S recycling by the HflX:control-apo-70S at 900ms Method: EM (single particle) / Resolution: 3.0 Å 29687 8g31 EMDB-29687, PDB-8g31: Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate Method: EM (single particle) / Resolution: 3.2 Å 29688 8g34 EMDB-29688, PDB-8g34: Time-resolved cryo-EM study of the 70S recycling by the HflX:1st intermediate Method: EM (single particle) / Resolution: 3.2 Å 29689 8g38 EMDB-29689, PDB-8g38: Time-resolved cryo-EM study of the 70S recycling by the HflX:3rd Intermediate Method: EM (single particle) / Resolution: 3.2 Å EMDB-29723: Time-resolved cryo-EM study of the 70S recycling by the HflX:3rd Intermediate, 30S focused map Method: EM (single particle) / Resolution: 4.1 Å EMDB-29724: Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate consensus, 30S focused Method: EM (single particle) / Resolution: 3.4 Å EMDB-29833: Time-resolved cryo-EM study of the 70S recycling by the HflX:1st intermediate, 30S focused Method: EM (single particle) / Resolution: 3.2 Å EMDB-29834: Time-resolved cryo-EM study of the 70S recycling by the HflX:1st intermediate, 50S focused from 30S subtracted Method: EM (single particle) / Resolution: 3.6 Å EMDB-29842: Time-resolved cryo-EM study of the 70S recycling by the HflX:3rd Intermediate, 50S focused and 30S subtracted Method: EM (single particle) / Resolution: 3.0 Å EMDB-29844: Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate, 50S focused and 30S subtracted Method: EM (single particle) / Resolution: 3.14 Å
Chemicals	ChemComp-GTP: GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying moleculeYM ChemComp-HOH*: WATER
Source	escherichia coli (E. coli)
Keywords	RIBOSOME / Recycling / Time-resolved Cryo-EM / 70S / HflX