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Yorodumi- PDB-8p8v: Mycoplasma pneumoniae di-ribosome in chloramphenicol-treated cell... -
+Open data
-Basic information
Entry | Database: PDB / ID: 8p8v | |||||||||
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Title | Mycoplasma pneumoniae di-ribosome in chloramphenicol-treated cells (leading 70S) | |||||||||
Components |
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Keywords | TRANSLATION / In situ / Ribosome / Chloramphenicol / cryo-ET | |||||||||
Function / homology | Function and homology information peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / large ribosomal subunit / protein transport / protein folding / regulation of translation / large ribosomal subunit rRNA binding / transferase activity / small ribosomal subunit / ribosomal large subunit assembly ...peptidylprolyl isomerase / peptidyl-prolyl cis-trans isomerase activity / large ribosomal subunit / protein transport / protein folding / regulation of translation / large ribosomal subunit rRNA binding / transferase activity / small ribosomal subunit / ribosomal large subunit assembly / cytosolic large ribosomal subunit / tRNA binding / rRNA binding / ribosome / structural constituent of ribosome / translation / ribonucleoprotein complex / cell division / response to antibiotic / mRNA binding / zinc ion binding / cytoplasm Similarity search - Function | |||||||||
Biological species | Mycoplasmoides pneumoniae M129 (bacteria) | |||||||||
Method | ELECTRON MICROSCOPY / subtomogram averaging / cryo EM / Resolution: 8.7 Å | |||||||||
Authors | Schacherl, M. / Xue, L. / Spahn, C.M.T. / Mahamid, J. | |||||||||
Funding support | United States, Germany, 2items
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Citation | Journal: Nat.Struct.Mol.Biol. / Year: 2024 Title: Structural insights into context-dependent inhibitory mechanisms of chloramphenicol in cells Authors: Xue, L. / Spahn, C.M.T. / Schacherl, M. / Mahamid, J. | |||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 8p8v.cif.gz | 3.6 MB | Display | PDBx/mmCIF format |
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PDB format | pdb8p8v.ent.gz | Display | PDB format | |
PDBx/mmJSON format | 8p8v.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8p8v_validation.pdf.gz | 1.8 MB | Display | wwPDB validaton report |
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Full document | 8p8v_full_validation.pdf.gz | 1.9 MB | Display | |
Data in XML | 8p8v_validation.xml.gz | 225.5 KB | Display | |
Data in CIF | 8p8v_validation.cif.gz | 392.4 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/p8/8p8v ftp://data.pdbj.org/pub/pdb/validation_reports/p8/8p8v | HTTPS FTP |
-Related structure data
Related structure data | 8p6pC 8p7xC 8p7yC 8p8bC 8p8wC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
Experimental dataset #1 | Data reference: 10.6019/EMPIAR-11520 / Data set type: EMPIAR |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
+50S ribosomal protein ... , 30 types, 30 molecules 0129abcdefghijklmnopqrstuvwxyz
-RNA chain , 7 types, 7 molecules 345678Y
#4: RNA chain | Mass: 940953.500 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: GenBank: 26117688 |
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#5: RNA chain | Mass: 34796.676 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: GenBank: 26117688 |
#6: RNA chain | Mass: 491327.844 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: GenBank: 26117688 |
#7: RNA chain | Mass: 24440.492 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: GenBank: 26117688 |
#8: RNA chain | Mass: 24136.244 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: GenBank: 26117688 |
#9: RNA chain | Mass: 24401.449 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: GenBank: 26117688 |
#32: RNA chain | Mass: 12936.888 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) |
-30S ribosomal protein ... , 20 types, 20 molecules ABCDEFGHIJKLMNOPQRST
#11: Protein | Mass: 33468.629 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75560 |
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#12: Protein | Mass: 30657.205 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P41205 |
#13: Protein | Mass: 23817.645 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P46775 |
#14: Protein | Mass: 24138.008 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: Q50301 |
#15: Protein | Mass: 25430.465 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75543 |
#16: Protein | Mass: 17897.029 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75545 |
#17: Protein | Mass: 15903.083 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: Q50304 |
#18: Protein | Mass: 15149.735 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75179 |
#19: Protein | Mass: 12226.571 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75581 |
#20: Protein | Mass: 12709.851 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: Q50296 |
#21: Protein | Mass: 15666.580 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75546 |
#22: Protein | Mass: 14209.607 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: Q50297 |
#23: Protein | Mass: 6901.364 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: Q50305 |
#24: Protein | Mass: 9921.687 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75173 |
#25: Protein | Mass: 10806.921 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: A0A0H3DLS7 |
#26: Protein | Mass: 9859.643 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: Q50309 |
#27: Protein | Mass: 12411.522 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75541 |
#28: Protein | Mass: 10057.626 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75576 |
#29: Protein | Mass: 9993.599 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75237 |
#30: Protein | Mass: 7539.182 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P57079 |
-Protein / Protein/peptide , 2 types, 2 molecules XZ
#31: Protein | Mass: 51427.031 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) / References: UniProt: P75454, peptidylprolyl isomerase |
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#33: Protein/peptide | Mass: 3127.794 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mycoplasmoides pneumoniae M129 (bacteria) |
-Non-polymers , 5 types, 331 molecules
#60: Chemical | ChemComp-ZN / #61: Chemical | ChemComp-CLM / | #62: Chemical | ChemComp-K / | #63: Chemical | ChemComp-MG / #64: Chemical | ChemComp-LYS / | |
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-Details
Has ligand of interest | Y |
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Has protein modification | Y |
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: CELL / 3D reconstruction method: subtomogram averaging |
-Sample preparation
Component | Name: Mycoplasma pneumoniae M129 cells treated with chloramphenicol Type: CELL / Details: Leading 70S in di-ribosome / Entity ID: #1-#8, #10-#59 / Source: NATURAL |
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Source (natural) | Organism: Mycoplasmoides pneumoniae M129 (bacteria) |
Buffer solution | pH: 7.4 Details: The modified Hayflick medium: 14.7g/L Difco PPLO(Becton Dickinson), 20% (v/v) Gibco horse serum(New Zealand origin), 100 mM HEPES-Na; pH 7.4, 1% (w/w) glucose, 0.002% (w/w) phenol red, 1000 U/mL penicillin G. |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: Mycoplasma pneumoniae M129 cells were grown on gold Quantifoil grids at 37 Celsius in the modified Hayflick medium. Treatment with chloramphenicol at the final concentration of 0.2 mg/ml was ...Details: Mycoplasma pneumoniae M129 cells were grown on gold Quantifoil grids at 37 Celsius in the modified Hayflick medium. Treatment with chloramphenicol at the final concentration of 0.2 mg/ml was performed for about 15 minutes before plunge freezing. |
Specimen support | Grid material: GOLD / Grid mesh size: 200 divisions/in. / Grid type: Quantifoil R2/1 |
Vitrification | Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE-PROPANE Details: Back-side blotting for 2-3 seconds before plunging using a manual plunger without an environmental control chamber. |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 64000 X / Nominal defocus max: 3250 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm / Alignment procedure: COMA FREE |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Electron dose: 3.34 e/Å2 / Avg electron dose per subtomogram: 137 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Num. of real images: 1 Details: Gatan K3 camera in non-CDS counting mode, targeted dose rate on camera ~20 e/pixel/second, 10 frames per tilt image, constant exposure time for each tilt, pixel size 1.329A |
EM imaging optics | Energyfilter name: GIF Bioquantum / Energyfilter slit width: 20 eV |
-Processing
EM software | Name: PHENIX / Version: 1.20.1_4487: / Category: model refinement | ||||||||||||||||||||||||
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Image processing | Details: 30774 sub-tomograms with large box size (3.1A/voxel pixel and box 256) were extracted, which can accommodate the neighbouring ribosomes(disomes). | ||||||||||||||||||||||||
CTF correction | Details: CTF estimation and 3D CTF correction are done in Warp Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||
3D reconstruction | Resolution: 8.7 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 963 / Algorithm: FOURIER SPACE / Details: half maps from RELION / Symmetry type: POINT | ||||||||||||||||||||||||
EM volume selection | Num. of tomograms: 139 / Num. of volumes extracted: 30774 | ||||||||||||||||||||||||
Atomic model building | Protocol: RIGID BODY FIT / Space: REAL / Target criteria: Cross-correlation coefficient | ||||||||||||||||||||||||
Atomic model building |
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Refine LS restraints |
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