[English] 日本語
Yorodumi- PDB-8a3d: Human mature large subunit of the ribosome with eIF6 and homoharr... -
+
Open data
-
Basic information
| Entry | Database: PDB / ID: 8a3d | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Title | Human mature large subunit of the ribosome with eIF6 and homoharringtonine bound | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Components |
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Keywords | TRANSLATION / Ribosome / human / rna modifications / water / magnesium / potassium / homoharringtonine | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Function / homology | Function and homology informationlamin filament / regulation of megakaryocyte differentiation / regulation of fatty acid biosynthetic process / miRNA-mediated post-transcriptional gene silencing / miRNA-mediated gene silencing by inhibition of translation / embryonic brain development / translation at presynapse / optic nerve development / alpha-beta T cell differentiation / exit from mitosis ...lamin filament / regulation of megakaryocyte differentiation / regulation of fatty acid biosynthetic process / miRNA-mediated post-transcriptional gene silencing / miRNA-mediated gene silencing by inhibition of translation / embryonic brain development / translation at presynapse / optic nerve development / alpha-beta T cell differentiation / exit from mitosis / regulation of translation involved in cellular response to UV / eukaryotic 80S initiation complex / ribosomal protein import into nucleus / regulation of G1 to G0 transition / axial mesoderm development / retinal ganglion cell axon guidance / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / regulation of reactive oxygen species metabolic process / middle ear morphogenesis / protein-DNA complex disassembly / 90S preribosome assembly / regulation of glycolytic process / negative regulation of formation of translation preinitiation complex / positive regulation of DNA damage response, signal transduction by p53 class mediator / GAIT complex / maturation of 5.8S rRNA / TORC2 complex binding / G1 to G0 transition / blastocyst development / negative regulation of myoblast fusion / cytoplasmic side of rough endoplasmic reticulum membrane / PELO:HBS1L and ABCE1 dissociate a ribosome on a non-stop mRNA / positive regulation of natural killer cell proliferation / ribosomal large subunit binding / homeostatic process / lung morphogenesis / ZNF598 and the Ribosome-associated Quality Trigger (RQT) complex dissociate a ribosome stalled on a no-go mRNA / macrophage chemotaxis / Protein hydroxylation / Peptide chain elongation / Selenocysteine synthesis / Formation of a pool of free 40S subunits / Eukaryotic Translation Termination / protein localization to nucleus / protein targeting / SRP-dependent cotranslational protein targeting to membrane / Response of EIF2AK4 (GCN2) to amino acid deficiency / ubiquitin ligase inhibitor activity / negative regulation of ubiquitin-dependent protein catabolic process / Viral mRNA Translation / retina development in camera-type eye / 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 / cellular response to interleukin-4 / L13a-mediated translational silencing of Ceruloplasmin expression / embryo implantation / 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 / Ribosome Quality Control (RQC) complex extracts and degrades nascent peptide / translation regulator activity / rough endoplasmic reticulum / translation initiation factor activity / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / ossification / MDM2/MDM4 family protein binding / regulation of signal transduction by p53 class mediator / skeletal system development / DNA damage response, signal transduction by p53 class mediator / cytosolic ribosome assembly / striated muscle contraction / ribosomal large subunit biogenesis / positive regulation of translation / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / assembly of large subunit precursor of preribosome / cytosolic ribosome / mRNA 3'-UTR binding / sensory perception of sound / innate immune response in mucosa / cellular response to type II interferon / cellular response to gamma radiation / response to insulin / bone development / mRNA 5'-UTR binding / transcription coactivator binding / Regulation of expression of SLITs and ROBOs / cytoplasmic ribonucleoprotein granule / cellular response to UV / rRNA processing / heparin binding / regulation of translation / antimicrobial humoral immune response mediated by antimicrobial peptide / large ribosomal subunit / antibacterial humoral response / presynapse / ribosome binding / 5S 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
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| History |
|
-
Structure visualization
| Structure viewer | Molecule: Molmil Jmol/JSmol |
|---|
-
Downloads & links
-
Download
| PDBx/mmCIF format | 8a3d.cif.gz | 3.7 MB | Display | PDBx/mmCIF format |
|---|---|---|---|---|
| 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 |
|---|
-Related structure data
| Related structure data | ![]() 15113MC M: map data used to model this data C: citing same article ( |
|---|---|
| Similar structure data | Similarity search - Function & homology F&H Search |
-
Links
-
Assembly
| Deposited unit | ![]()
|
|---|---|
| 1 |
|
-
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 |
|---|---|
| #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 |
|---|---|
| #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 / | |
|---|
-Details
| Has ligand of interest | N |
|---|---|
| Has protein modification | N |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
|---|---|
| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-
Sample preparation
| Component | Name: Human mature large subunit of the ribosome with eIF6 bound Type: RIBOSOME / Entity ID: #1-#45 / Source: NATURAL | |||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Molecular weight | Experimental value: NO | |||||||||||||||||||||||||
| Source (natural) | Organism: Homo sapiens (human) | |||||||||||||||||||||||||
| Buffer solution | pH: 7.5 | |||||||||||||||||||||||||
| Buffer component |
| |||||||||||||||||||||||||
| 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 |
-
Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
|---|---|
| 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 |
-
Processing
| Software | Name: REFMAC / Version: 5.8.0267 / Classification: refinement | ||||||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| EM software |
| ||||||||||||||||||||||||||||||||||||
| 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
| ||||||||||||||||||||||||||||||||||||
| Displacement parameters | Biso mean: 35.442 Å2 | ||||||||||||||||||||||||||||||||||||
| LS refinement shell | Highest resolution: 1.67 Å /
|
Movie
Controller
About Yorodumi



Homo sapiens (human)
United Kingdom, 1items
Citation
PDBj






































FIELD EMISSION GUN
