8G31
Time-resolved cryo-EM study of the 70S recycling by the HflX:2nd Intermediate
This is a non-PDB format compatible entry.
Summary for 8G31
| Entry DOI | 10.2210/pdb8g31/pdb |
| EMDB information | 29681 29687 29688 29689 29724 29844 |
| Descriptor | 50S ribosomal protein L32, 50S ribosomal protein L3, 50S ribosomal protein L4, ... (52 entities in total) |
| Functional Keywords | recycling, time-resolved cryo-em, 70s, hflx, ribosome |
| Biological source | Escherichia coli More |
| Total number of polymer chains | 52 |
| Total formula weight | 2140230.47 |
| Authors | Bhattacharjee, S.,Brown, P.Z.,Frank, J. (deposition date: 2023-02-06, release date: 2023-12-06, Last modification date: 2024-02-14) |
| Primary citation | 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 Cited by 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: 38244547DOI: 10.1016/j.cell.2023.12.027 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.2 Å) |
Structure validation
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