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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 / eukaryotic 80S initiation complex / ribosomal protein import into nucleus / regulation of G1 to G0 transition ...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 / eukaryotic 80S initiation complex / ribosomal protein import into nucleus / regulation of G1 to G0 transition / regulation of reactive oxygen species metabolic process / regulation of glycolytic process / G1 to G0 transition / negative regulation of formation of translation preinitiation complex / GAIT complex / maturation of 5.8S rRNA / TORC2 complex binding / cytoplasmic side of rough endoplasmic reticulum membrane / regulation of translation involved in cellular response to UV / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / negative regulation of myoblast fusion / protein-DNA complex disassembly / ribosomal large subunit binding / positive regulation of DNA damage response, signal transduction by p53 class mediator / Protein hydroxylation / PELO:HBS1L and ABCE1 dissociate a ribosome on a non-stop mRNA / protein localization to nucleus / Peptide chain elongation / protein targeting / Selenocysteine synthesis / Formation of a pool of free 40S subunits / Eukaryotic Translation Termination / SRP-dependent cotranslational protein targeting to membrane / Response of EIF2AK4 (GCN2) to amino acid deficiency / ubiquitin ligase inhibitor activity / Viral mRNA Translation / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / positive regulation of signal transduction by p53 class mediator / ribosomal subunit export from nucleus / GTP hydrolysis and joining of the 60S ribosomal subunit / embryo implantation / L13a-mediated translational silencing of Ceruloplasmin expression / Major pathway of rRNA processing in the nucleolus and cytosol / maturation of LSU-rRNA / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / protein-RNA complex assembly / negative regulation of ubiquitin-dependent protein catabolic process / translation regulator activity / rough endoplasmic reticulum / translation initiation factor activity / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / MDM2/MDM4 family protein binding / regulation of signal transduction by p53 class mediator / cytosolic ribosome assembly / ZNF598 and the Ribosome-associated Quality Trigger (RQT) complex dissociate a ribosome stalled on a no-go mRNA / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / DNA damage response, signal transduction by p53 class mediator / assembly of large subunit precursor of preribosome / striated muscle contraction / mRNA 3'-UTR binding / positive regulation of translation / cellular response to type II interferon / response to insulin / bone development / cellular response to gamma radiation / fibrillar center / cytoplasmic ribonucleoprotein granule / Regulation of expression of SLITs and ROBOs / mRNA 5'-UTR binding / innate immune response in mucosa / transcription coactivator binding / Ribosome Quality Control (RQC) complex extracts and degrades nascent peptide / cellular response to UV / rRNA processing / cytosolic ribosome / heparin binding / presynapse / large ribosomal subunit / ribosome binding / Dengue Virus-Host Interactions / 5S rRNA binding / ribosomal large subunit assembly / cell body / antimicrobial humoral immune response mediated by antimicrobial peptide / large ribosomal subunit rRNA binding / spermatogenesis / antibacterial humoral response / killing of cells of another organism / defense response to Gram-negative bacterium / cytosolic large ribosomal subunit / tRNA binding / cytoplasmic translation / postsynapse / defense response to Gram-positive bacterium / postsynaptic density / intracellular signal transduction / negative regulation of translation / protein stabilization / rRNA binding 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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| Has protein modification | N |
-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
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FIELD EMISSION GUN
