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Yorodumi- PDB-7bge: Staphylococcus aureus 30S ribosomal subunit in presence of spermi... -
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Basic information
| Entry | Database: PDB / ID: 7bge | |||||||||
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| Title | Staphylococcus aureus 30S ribosomal subunit in presence of spermidine (head only) | |||||||||
Components |
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Keywords | RIBOSOME / Pathogen / small ribosomal subunit / spermidine | |||||||||
| Function / homology | Function and homology informationribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / tRNA binding / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / mRNA binding ...ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / tRNA binding / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / mRNA binding / RNA binding / zinc ion binding / cytoplasm / cytosol Similarity search - Function | |||||||||
| Biological species | Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria)![]() | |||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||
Authors | Belinite, M. / Khusainov, I. / Marzi, S. / Romby, P. / Yusupov, M. / Hashem, Y. | |||||||||
| Funding support | France, 2items
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Citation | Journal: Front Mol Biosci / Year: 2021Title: 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. | |||||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 7bge.cif.gz | 416.4 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb7bge.ent.gz | 302.6 KB | Display | PDB format |
| PDBx/mmJSON format | 7bge.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 7bge_validation.pdf.gz | 773.5 KB | Display | wwPDB validaton report |
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| Full document | 7bge_full_validation.pdf.gz | 835.2 KB | Display | |
| Data in XML | 7bge_validation.xml.gz | 39.3 KB | Display | |
| Data in CIF | 7bge_validation.cif.gz | 64.1 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/bg/7bge ftp://data.pdbj.org/pub/pdb/validation_reports/bg/7bge | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 12179MC ![]() 7bgdC ![]() 7kwgC M: map data used to model this data C: citing same article ( |
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| Similar structure data |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-RNA chain , 1 types, 1 molecules a
| #1: RNA chain | Mass: 503218.031 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria) |
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-30S ribosomal protein ... , 8 types, 8 molecules bcgijmns
| #2: Protein | Mass: 29136.369 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus (strain NCTC 8325) (bacteria)Strain: NCTC 8325 / References: UniProt: Q2FZ25 |
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| #3: Protein | Mass: 24143.867 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus (strain NCTC 8325) (bacteria)Strain: NCTC 8325 / References: UniProt: Q2FW12 |
| #4: Protein | Mass: 17826.555 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus (strain NCTC 8325) (bacteria)Strain: NCTC 8325 / References: UniProt: P48940 |
| #5: Protein | Mass: 14856.987 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus (strain NCTC 8325) (bacteria)Strain: NCTC 8325 / References: UniProt: Q2FW39 |
| #6: Protein | Mass: 11600.520 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria)References: UniProt: Q931G5 |
| #7: Protein | Mass: 13747.919 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus (strain NCTC 8325) (bacteria)Strain: NCTC 8325 / References: UniProt: Q2FW30 |
| #8: Protein | Mass: 7317.769 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus (strain NCTC 8325) (bacteria)Strain: NCTC 8325 / References: UniProt: Q2FW19 |
| #9: Protein | Mass: 10639.309 Da / Num. of mol.: 1 / Source method: isolated from a natural source Source: (natural) Staphylococcus aureus (strain NCTC 8325) (bacteria)Strain: NCTC 8325 / References: UniProt: Q2FW10 |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
| Component | Name: Staphylococcus aureus 30S ribosomal subunit in presence of spermidine (head only) Type: RIBOSOME / Entity ID: all / Source: NATURAL |
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| Source (natural) | Organism: Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria) |
| Buffer solution | pH: 7.5 |
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TALOS ARCTICA |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: OTHER |
| Electron lens | Mode: BRIGHT FIELD |
| Image recording | Electron dose: 3 e/Å2 / Film or detector model: FEI FALCON III (4k x 4k) |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||
| 3D reconstruction | Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 529602 / Symmetry type: POINT |
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Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria)
France, 2items
Citation
UCSF Chimera















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