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Yorodumi- PDB-8a3d: Human mature large subunit of the ribosome with eIF6 and homoharr... -
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Open data
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Basic information
| Entry | Database: PDB / ID: 8a3d | ||||||
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| Title | Human mature large subunit of the ribosome with eIF6 and homoharringtonine bound | ||||||
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Keywords | TRANSLATION / Ribosome / human / rna modifications / water / magnesium / potassium / homoharringtonine | ||||||
| Function / homology | Function and homology informationlamin filament / regulation of fatty acid biosynthetic process / regulation of megakaryocyte differentiation / miRNA-mediated post-transcriptional gene silencing / miRNA-mediated gene silencing by inhibition of translation / embryonic brain development / translation at presynapse / exit from mitosis / optic nerve development / eukaryotic 80S initiation complex ...lamin filament / regulation of fatty acid biosynthetic process / regulation of megakaryocyte differentiation / miRNA-mediated post-transcriptional gene silencing / miRNA-mediated gene silencing by inhibition of translation / embryonic brain development / translation at presynapse / exit from mitosis / optic nerve development / eukaryotic 80S initiation complex / regulation of translation involved in cellular response to UV / negative regulation of protein neddylation / negative regulation of formation of translation preinitiation complex / axial mesoderm development / regulation of G1 to G0 transition / retinal ganglion cell axon guidance / ribosomal protein import into nucleus / protein-DNA complex disassembly / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / 90S preribosome assembly / alpha-beta T cell differentiation / GAIT complex / regulation of glycolytic process / positive regulation of DNA damage response, signal transduction by p53 class mediator / regulation of reactive oxygen species metabolic process / TORC2 complex binding / G1 to G0 transition / maturation of 5.8S rRNA / middle ear morphogenesis / cytoplasmic side of rough endoplasmic reticulum membrane / negative regulation of ubiquitin protein ligase activity / homeostatic process / ribosomal large subunit binding / macrophage chemotaxis / lung morphogenesis / cellular response to actinomycin D / positive regulation of natural killer cell proliferation / Protein hydroxylation / Peptide chain elongation / positive regulation of protein binding / Selenocysteine synthesis / Formation of a pool of free 40S subunits / Eukaryotic Translation Termination / blastocyst development / SRP-dependent cotranslational protein targeting to membrane / Response of EIF2AK4 (GCN2) to amino acid deficiency / negative regulation of ubiquitin-dependent protein catabolic process / ubiquitin ligase inhibitor activity / Viral mRNA Translation / positive regulation of signal transduction by p53 class mediator / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / protein localization to nucleus / GTP hydrolysis and joining of the 60S ribosomal subunit / L13a-mediated translational silencing of Ceruloplasmin expression / Major pathway of rRNA processing in the nucleolus and cytosol / protein targeting / ribosomal subunit export from nucleus / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / protein-RNA complex assembly / maturation of LSU-rRNA / rough endoplasmic reticulum / translation regulator activity / translation initiation factor activity / MDM2/MDM4 family protein binding / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / embryo implantation / cellular response to interleukin-4 / cytosolic ribosome / ossification / assembly of large subunit precursor of preribosome / regulation of signal transduction by p53 class mediator / cytosolic ribosome assembly / positive regulation of translation / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / skeletal system development / innate immune response in mucosa / mRNA 3'-UTR binding / DNA damage response, signal transduction by p53 class mediator / sensory perception of sound / cellular response to gamma radiation / bone development / response to insulin / mRNA 5'-UTR binding / cellular response to type II interferon / transcription coactivator binding / Regulation of expression of SLITs and ROBOs / cytoplasmic ribonucleoprotein granule / rRNA processing / cellular response to UV / antimicrobial humoral immune response mediated by antimicrobial peptide / heparin binding / regulation of translation / large ribosomal subunit / antibacterial humoral response / presynapse / ribosome binding / retina development in camera-type eye / cell body / ribosomal large subunit assembly Similarity search - Function | ||||||
| Biological species | Homo sapiens (human) | ||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 1.67 Å | ||||||
Authors | Faille, A. / Warren, A.J. / Dent, K.C. | ||||||
| Funding support | United Kingdom, 1items
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Citation | Journal: bioRxiv / Year: 2023Title: The chemical landscape of the human ribosome at 1.67 Å resolution. Authors: Alexandre Faille / Kyle C Dent / Simone Pellegrino / Pekka Jaako / Alan J Warren Abstract: The ability of ribosomes to translate the genetic code into protein requires a finely tuned ion and solvent ecosystem. However, the lack of high-resolution structures has precluded accurate ...The ability of ribosomes to translate the genetic code into protein requires a finely tuned ion and solvent ecosystem. However, the lack of high-resolution structures has precluded accurate positioning of all the functional elements of the ribosome and limited our understanding of the specific role of ribosomal RNA chemical modifications in modulating ribosome function in health and disease. Here, using a new sample preparation methodology based on functionalised pristine graphene-coated grids, we solve the cryo-EM structure of the human large ribosomal subunit to a resolution of 1.67 Å. The accurate assignment of water molecules, magnesium and potassium ions in our model highlights the fundamental biological role of ribosomal RNA methylation in harnessing unconventional carbon-oxygen hydrogen bonds to establish chemical interactions with the environment and fine-tune the functional interplay with tRNA. In addition, the structures of three translational inhibitors bound to the human large ribosomal subunit at better than 2 Å resolution provide mechanistic insights into how three key druggable pockets of the ribosome are targeted and illustrate the potential of this methodology to accelerate high-throughput structure-based design of anti-cancer therapeutics. | ||||||
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Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 8a3d.cif.gz | 3.7 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb8a3d.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 8a3d.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/a3/8a3d ftp://data.pdbj.org/pub/pdb/validation_reports/a3/8a3d | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 15113MC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-RNA chain , 3 types, 3 molecules ABC
| #1: RNA chain | Mass: 1640812.500 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Cell line: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / References: GenBank: 86475748 |
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| #2: RNA chain | Mass: 38998.078 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Cell line: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / References: GenBank: 23898 |
| #3: RNA chain | Mass: 50463.840 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Cell line: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / References: GenBank: 555853 |
+60S ribosomal protein ... , 39 types, 39 molecules DEFGHmnopqrslIJKLMNOPQRSTUVWXY...
-Protein , 3 types, 3 molecules bgh
| #36: Protein | Mass: 21730.748 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Cell line: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 |
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| #41: Protein | Mass: 11470.713 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Cell line: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / References: UniProt: A0A3Q0DRJ6 |
| #45: Protein | Mass: 26620.010 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / Cell line: HEK293 / Plasmid details: Thermofisher / Variant: Expi293 / References: UniProt: P56537 |
-Non-polymers , 8 types, 12367 molecules 














| #46: Chemical | ChemComp-MG / #47: Chemical | ChemComp-K / #48: Chemical | #49: Chemical | ChemComp-HMT / ( | #50: Chemical | ChemComp-SPM / | #51: Chemical | ChemComp-EPE / | #52: Chemical | ChemComp-ZN / #53: Water | ChemComp-HOH / | |
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-Details
| Has ligand of interest | N |
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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 |
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Sample preparation
| Component | Name: Human mature large subunit of the ribosome with eIF6 bound Type: RIBOSOME / Entity ID: #1-#45 / Source: NATURAL | |||||||||||||||||||||||||
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| Molecular weight | Experimental value: NO | |||||||||||||||||||||||||
| Source (natural) | Organism: Homo sapiens (human) | |||||||||||||||||||||||||
| Buffer solution | pH: 7.5 | |||||||||||||||||||||||||
| Buffer component |
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| Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | |||||||||||||||||||||||||
| Specimen support | Details: Grid was functionalised with 1-pyrenemethylamine. See methods section in published article. Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R2/2 | |||||||||||||||||||||||||
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K / Details: 30 seconds 'wait' time |
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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: 105000 X / Nominal defocus max: 1800 nm / Nominal defocus min: 1000 nm / Cs: 2.7 mm / C2 aperture diameter: 50 µm / Alignment procedure: COMA FREE |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Average exposure time: 2.2 sec. / Electron dose: 51 e/Å2 / Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 9973 |
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Processing
| Software | Name: REFMAC / Version: 5.8.0267 / Classification: refinement | ||||||||||||||||||||||||||||||||||||
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| EM software |
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 1147785 | ||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 1.67 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 1088709 / Num. of class averages: 1 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||
| Atomic model building | B value: 40 / Protocol: AB INITIO MODEL / Space: REAL | ||||||||||||||||||||||||||||||||||||
| Atomic model building | PDB-ID: 6EK0![]() 6ek0 Accession code: 6EK0 / Source name: PDB / Type: experimental model | ||||||||||||||||||||||||||||||||||||
| Refinement | Resolution: 1.67→270.25 Å / Cor.coef. Fo:Fc: 0.688
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| Displacement parameters | Biso mean: 35.442 Å2 | ||||||||||||||||||||||||||||||||||||
| LS refinement shell | Highest resolution: 1.67 Å /
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Homo sapiens (human)
United Kingdom, 1items
Citation
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FIELD EMISSION GUN
