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Yorodumi- EMDB-12091: Staphylococcus aureus 30S ribosomal subunit in absence of spermid... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-12091 | |||||||||
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Title | Staphylococcus aureus 30S ribosomal subunit in absence of spermidine with reoriented helix 44 | |||||||||
Map data | ||||||||||
Sample |
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Function / homology | Function and homology information ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation ...ribosomal small subunit biogenesis / small ribosomal subunit rRNA binding / ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / mRNA binding / RNA binding / zinc ion binding / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 13.76 Å | |||||||||
Authors | Belinite M / Khusainov I / Marzi S / Romby P / Yusupov M / Hashem Y | |||||||||
Funding support | France, 2 items
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Citation | Journal: Front Mol Biosci / Year: 2021 Title: Stabilization of Ribosomal RNA of the Small Subunit by Spermidine in . Authors: Margarita Belinite / Iskander Khusainov / Heddy Soufari / Stefano Marzi / Pascale Romby / Marat Yusupov / Yaser Hashem / Abstract: Cryo-electron microscopy is now used as a method of choice in structural biology for studying protein synthesis, a process mediated by the ribosome machinery. In order to achieve high-resolution ...Cryo-electron microscopy is now used as a method of choice in structural biology for studying protein synthesis, a process mediated by the ribosome machinery. In order to achieve high-resolution structures using this approach, one needs to obtain homogeneous and stable samples, which requires optimization of ribosome purification in a species-dependent manner. This is especially critical for the bacterial small ribosomal subunit that tends to be unstable in the absence of ligands. Here, we report a protocol for purification of stable 30 S from the Gram-positive bacterium and its cryo-EM structures: in presence of spermidine at a resolution ranging between 3.4 and 3.6 Å and in its absence at 5.3 Å. Using biochemical characterization and cryo-EM, we demonstrate the importance of spermidine for stabilization of the 30 S preserving favorable conformation of the helix 44. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_12091.map.gz | 618.4 KB | EMDB map data format | |
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Header (meta data) | emd-12091-v30.xml emd-12091.xml | 8.2 KB 8.2 KB | Display Display | EMDB header |
Images | emd_12091.png | 60.8 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12091 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12091 | HTTPS FTP |
-Validation report
Summary document | emd_12091_validation.pdf.gz | 314.9 KB | Display | EMDB validaton report |
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Full document | emd_12091_full_validation.pdf.gz | 314.5 KB | Display | |
Data in XML | emd_12091_validation.xml.gz | 4.8 KB | Display | |
Data in CIF | emd_12091_validation.cif.gz | 5.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12091 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12091 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_12091.map.gz / Format: CCP4 / Size: 931.6 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 4.96 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : Staphylococcus aureus 30S ribosomal subunit in absence of spermid...
Entire | Name: Staphylococcus aureus 30S ribosomal subunit in absence of spermidine, class 5 |
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Components |
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-Supramolecule #1: Staphylococcus aureus 30S ribosomal subunit in absence of spermid...
Supramolecule | Name: Staphylococcus aureus 30S ribosomal subunit in absence of spermidine, class 5 type: complex / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: FEI FALCON III (4k x 4k) / Average electron dose: 3.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
-Image processing
CTF correction | Software - Name: Gctf (ver. 1.06) |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 13.76 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 215450 |
Initial angle assignment | Type: PROJECTION MATCHING |
Final angle assignment | Type: PROJECTION MATCHING |