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Yorodumi- PDB-7bge: Staphylococcus aureus 30S ribosomal subunit in presence of spermi... -
+Open data
-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 information ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / mRNA binding / RNA binding ...ribosomal small subunit assembly / small ribosomal subunit / cytosolic small ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / mRNA binding / RNA binding / zinc ion binding / cytosol Similarity search - Function | |||||||||
Biological species | Staphylococcus aureus subsp. aureus NCTC 8325 (bacteria) Staphylococcus aureus (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: 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 | Molecule: MolmilJmol/JSmol |
-Downloads & links
-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 (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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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 |
-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 |
-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) |
-Processing
EM software |
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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 |