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Yorodumi- PDB-4v6z: E. coli 70S-fMetVal-tRNAVal-tRNAfMet complex in classic pre-trans... -
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Basic information
| Entry | Database: PDB / ID: 4v6z | ||||||||||||
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| Title | E. coli 70S-fMetVal-tRNAVal-tRNAfMet complex in classic pre-translocation state (pre1b) | ||||||||||||
|  Components | 
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|  Keywords | RIBOSOME / cryo-EM refinement / tRNA / translocation intermediate | ||||||||||||
| Function / homology |  Function and homology information stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / positive regulation of ribosome biogenesis / RNA-binding transcription regulator activity ...stringent response / ornithine decarboxylase inhibitor activity / transcription antitermination factor activity, RNA binding / misfolded RNA binding / Group I intron splicing / RNA folding / transcriptional attenuation / endoribonuclease inhibitor activity / positive regulation of ribosome biogenesis / RNA-binding transcription regulator activity / translational termination / negative regulation of cytoplasmic translation / four-way junction DNA binding / DnaA-L2 complex / translation repressor activity / negative regulation of translational initiation / regulation of mRNA stability / negative regulation of DNA-templated DNA replication initiation / mRNA regulatory element binding translation repressor activity / assembly of large subunit precursor of preribosome / positive regulation of RNA splicing / ribosome assembly / regulation of DNA-templated transcription elongation / transcription elongation factor complex / cytosolic ribosome assembly / response to reactive oxygen species / DNA endonuclease activity / transcription antitermination / regulation of cell growth / DNA-templated transcription termination / response to radiation / maintenance of translational fidelity / mRNA 5'-UTR binding / ribosome biogenesis / large ribosomal subunit / regulation of translation / ribosome binding / transferase activity / ribosomal small subunit biogenesis / ribosomal small subunit assembly / small ribosomal subunit / 5S rRNA binding / small ribosomal subunit rRNA binding / ribosomal large subunit assembly / cytosolic small ribosomal subunit / large ribosomal subunit rRNA binding / cytosolic large ribosomal subunit / cytoplasmic translation / tRNA binding / negative regulation of translation / rRNA binding / structural constituent of ribosome / ribosome / translation / response to antibiotic / negative regulation of DNA-templated transcription / mRNA binding / DNA binding / RNA binding / zinc ion binding / membrane / cytoplasm / cytosol Similarity search - Function | ||||||||||||
| Biological species |   Escherichia coli (E. coli) | ||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 12 Å | ||||||||||||
|  Authors | Blau, C. / Bock, L.V. / Schroder, G.F. / Davydov, I. / Fischer, N. / Stark, H. / Rodnina, M.V. / Vaiana, A.C. / Grubmuller, H. | ||||||||||||
|  Citation |  Journal: Nat Struct Mol Biol / Year: 2013 Title: Energy barriers and driving forces in tRNA translocation through the ribosome. Authors: Lars V Bock / Christian Blau / Gunnar F Schröder / Iakov I Davydov / Niels Fischer / Holger Stark / Marina V Rodnina / Andrea C Vaiana / Helmut Grubmüller /  Abstract: During protein synthesis, tRNAs move from the ribosome's aminoacyl to peptidyl to exit sites. Here we investigate conformational motions during spontaneous translocation, using molecular dynamics ...During protein synthesis, tRNAs move from the ribosome's aminoacyl to peptidyl to exit sites. Here we investigate conformational motions during spontaneous translocation, using molecular dynamics simulations of 13 intermediate-translocation-state models obtained by combining Escherichia coli ribosome crystal structures with cryo-EM data. Resolving fast transitions between states, we find that tRNA motions govern the transition rates within the pre- and post-translocation states. Intersubunit rotations and L1-stalk motion exhibit fast intrinsic submicrosecond dynamics. The L1 stalk drives the tRNA from the peptidyl site and links intersubunit rotation to translocation. Displacement of tRNAs is controlled by 'sliding' and 'stepping' mechanisms involving conserved L16, L5 and L1 residues, thus ensuring binding to the ribosome despite large-scale tRNA movement. Our results complement structural data with a time axis, intrinsic transition rates and molecular forces, revealing correlated functional motions inaccessible by other means. | ||||||||||||
| History | 
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| Structure viewer | Molecule:  Molmil  Jmol/JSmol | 
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| PDBx/mmCIF format |  4v6z.cif.gz | 3.3 MB | Display |  PDBx/mmCIF format | 
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| PDB format |  pdb4v6z.ent.gz | Display |  PDB format | |
| PDBx/mmJSON format |  4v6z.json.gz | Tree view |  PDBx/mmJSON format | |
| Others |  Other downloads | 
-Validation report
| Summary document |  4v6z_validation.pdf.gz | 1.4 MB | Display |  wwPDB validaton report | 
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| Full document |  4v6z_full_validation.pdf.gz | 2.6 MB | Display | |
| Data in XML |  4v6z_validation.xml.gz | 306.9 KB | Display | |
| Data in CIF |  4v6z_validation.cif.gz | 502 KB | Display | |
| Arichive directory |  https://data.pdbj.org/pub/pdb/validation_reports/v6/4v6z  ftp://data.pdbj.org/pub/pdb/validation_reports/v6/4v6z | HTTPS FTP | 
-Related structure data
| Related structure data |  2472MC  2473C  2474C  2475C  4v6yC  4v70C  4v71C  4v72C  4v73C  4v74C  4v75C  4v76C  4v77C  4v78C  4v79C  4v7aC M: map data used to model this data C: citing same article ( | 
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| Similar structure data | 
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Links
- Assembly
Assembly
| Deposited unit |  
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| 1 | 
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- Components
Components
-30S ribosomal protein  ... , 20 types, 20 molecules ABACADAEAFAGAHAIAJAKALAMANAOAPAQARASATAU                   
| #1: Protein | Mass: 24277.992 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7V0 | 
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| #2: Protein | Mass: 23207.990 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7V3 | 
| #3: Protein | Mass: 23514.199 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7V8 | 
| #4: Protein | Mass: 15828.328 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7W1 | 
| #5: Protein | Mass: 11667.380 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P02358 | 
| #6: Protein | Mass: 16788.453 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P02359 | 
| #7: Protein | Mass: 14146.557 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7W7 | 
| #8: Protein | Mass: 14580.919 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7X3 | 
| #9: Protein | Mass: 11221.034 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7R5 | 
| #10: Protein | Mass: 12513.237 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7R9 | 
| #11: Protein | Mass: 13768.157 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7S3 | 
| #12: Protein | Mass: 12657.843 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7S9 | 
| #13: Protein | Mass: 11606.560 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0AG59 | 
| #14: Protein | Mass: 10290.816 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0ADZ4 | 
| #15: Protein | Mass: 9063.426 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7T3 | 
| #16: Protein | Mass: 9287.992 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0AG63 | 
| #17: Protein | Mass: 6490.523 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7T7 | 
| #18: Protein | Mass: 9081.672 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7U3 | 
| #19: Protein | Mass: 9532.228 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P0A7U7 | 
| #20: Protein | Mass: 6091.125 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 / References: UniProt: P68679 | 
-RNA chain , 6 types, 6 molecules AAA1A2A3BABB     
| #21: RNA chain | Mass: 496892.375 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 | 
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| #22: RNA chain | Mass: 24617.768 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 | 
| #23: RNA chain | Mass: 4693.746 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: mRNA | 
| #24: RNA chain | Mass: 24848.918 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 | 
| #54: RNA chain | Mass: 941306.188 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 | 
| #55: RNA chain | Mass: 38177.762 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural)    Escherichia coli (E. coli) / Strain: K12 | 
+50S ribosomal protein  ... , 30 types, 30 molecules BCBDBEBFBGBHBIBJBKBLBMBNBOBPBQBRBSBTBUBVBWBXBYBZB0B1B2B3B4B5                             
-Non-polymers , 2 types, 2 molecules 


| #57: Chemical | ChemComp-VAL / | 
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| #58: Chemical | ChemComp-FME / | 
-Details
| Has protein modification | Y | 
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-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY | 
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction | 
- Sample preparation
Sample preparation
| Component | Name: E. coli 70S-fMetVal-tRNAVal-tRNAfMet complex / Type: RIBOSOME | 
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| Buffer solution | pH: 7.5 | 
| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | 
| Vitrification | Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE | 
- Electron microscopy imaging
Electron microscopy imaging
| Microscopy | Model: FEI/PHILIPS CM200FEG / Date: May 11, 2008 | 
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| Electron gun | Electron source:  FIELD EMISSION GUN / Accelerating voltage: 160 kV / Illumination mode: SPOT SCAN | 
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 161000 X / Nominal defocus max: 2000 nm / Nominal defocus min: 500 nm | 
| Image recording | Electron dose: 20 e/Å2 / Film or detector model: GENERIC TVIPS (4k x 4k) | 
- Processing
Processing
| EM software | 
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| Symmetry | Point symmetry: C1 (asymmetric) | |||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 12 Å / Resolution method: FSC 0.5 CUT-OFF / Num. of particles: 13091 / Nominal pixel size: 2.8 Å / Actual pixel size: 2.8 Å / Symmetry type: POINT | |||||||||||||||||||||||||||||||||||
| Atomic model building | Protocol: FLEXIBLE FIT / Space: REAL / Target criteria: Cross-correlation coefficient Details: METHOD--Cross-correlation gradient based REFINEMENT PROTOCOL--flexible fitting, dynamic elastic network | |||||||||||||||||||||||||||||||||||
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