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Open data
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Basic information
| Entry | Database: PDB / ID: 6w6l | ||||||||||||||||||
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| Title | Cryo-EM structure of the human ribosome-TMCO1 translocon | ||||||||||||||||||
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Keywords | RIBOSOME/PROTEIN TRANSPORT / RIBOSOME-PROTEIN TRANSPORT complex | ||||||||||||||||||
| Function / homology | Function and homology informationnegative regulation of nodal signaling pathway / multi-pass transmembrane protein insertion into ER membrane / multi-pass translocon complex / endoplasmic reticulum Sec complex / determination of left/right asymmetry in lateral mesoderm / pronephric nephron development / rough endoplasmic reticulum membrane / cotranslational protein targeting to membrane / endoplasmic reticulum quality control compartment / Ssh1 translocon complex ...negative regulation of nodal signaling pathway / multi-pass transmembrane protein insertion into ER membrane / multi-pass translocon complex / endoplasmic reticulum Sec complex / determination of left/right asymmetry in lateral mesoderm / pronephric nephron development / rough endoplasmic reticulum membrane / cotranslational protein targeting to membrane / endoplasmic reticulum quality control compartment / Ssh1 translocon complex / Sec61 translocon complex / protein localization to nuclear inner membrane / protein insertion into ER membrane / protein targeting to ER / post-translational protein targeting to endoplasmic reticulum membrane / regulation of protein complex stability / SRP-dependent cotranslational protein targeting to membrane, translocation / embryonic brain development / signal sequence binding / SRP-dependent cotranslational protein targeting to membrane / translation at presynapse / post-translational protein targeting to membrane, translocation / endoplasmic reticulum calcium ion homeostasis / exit from mitosis / optic nerve development / endoplasmic reticulum organization / 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 / retinal ganglion cell axon guidance / 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 / positive regulation of DNA damage response, signal transduction by p53 class mediator / epidermal growth factor binding / TORC2 complex binding / retrograde protein transport, ER to cytosol / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / middle ear morphogenesis / ER overload response / cytoplasmic side of rough endoplasmic reticulum membrane / negative regulation of ubiquitin protein ligase activity / homeostatic process / macrophage chemotaxis / lung morphogenesis / positive regulation of natural killer cell proliferation / Protein hydroxylation / Peptide chain elongation / positive regulation of protein binding / protein transmembrane transporter activity / Selenocysteine synthesis / Formation of a pool of free 40S subunits / protein folding chaperone complex / Eukaryotic Translation Termination / blastocyst development / regulation of signal transduction / SRP-dependent cotranslational protein targeting to membrane / Response of EIF2AK4 (GCN2) to amino acid deficiency / negative regulation of ubiquitin-dependent protein catabolic process / response to type II interferon / regulation of protein-containing complex assembly / 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 / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / protein-RNA complex assembly / maturation of LSU-rRNA / ERAD pathway / rough endoplasmic reticulum / translation regulator activity / protein folding chaperone / MDM2/MDM4 family protein binding / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / embryo implantation / cellular response to interleukin-4 / cytosolic ribosome / ossification / guanyl-nucleotide exchange factor activity / regulation of signal transduction by p53 class mediator / positive regulation of translation / ribosomal large subunit biogenesis / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / post-embryonic development / 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 / mRNA 5'-UTR binding Similarity search - Function | ||||||||||||||||||
| Biological species | Homo sapiens (human) | ||||||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.84 Å | ||||||||||||||||||
Authors | Keenan, R.J. / McGilvray, P.T. | ||||||||||||||||||
| Funding support | United States, 5items
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Citation | Journal: Elife / Year: 2020Title: An ER translocon for multi-pass membrane protein biogenesis. Authors: Philip T McGilvray / S Andrei Anghel / Arunkumar Sundaram / Frank Zhong / Michael J Trnka / James R Fuller / Hong Hu / Alma L Burlingame / Robert J Keenan / ![]() Abstract: Membrane proteins with multiple transmembrane domains play critical roles in cell physiology, but little is known about the machinery coordinating their biogenesis at the endoplasmic reticulum. Here ...Membrane proteins with multiple transmembrane domains play critical roles in cell physiology, but little is known about the machinery coordinating their biogenesis at the endoplasmic reticulum. Here we describe a ~ 360 kDa ribosome-associated complex comprising the core Sec61 channel and five accessory factors: TMCO1, CCDC47 and the Nicalin-TMEM147-NOMO complex. Cryo-electron microscopy reveals a large assembly at the ribosome exit tunnel organized around a central membrane cavity. Similar to protein-conducting channels that facilitate movement of transmembrane segments, cytosolic and luminal funnels in TMCO1 and TMEM147, respectively, suggest routes into the central membrane cavity. High-throughput mRNA sequencing shows selective translocon engagement with hundreds of different multi-pass membrane proteins. Consistent with a role in multi-pass membrane protein biogenesis, cells lacking different accessory components show reduced levels of one such client, the glutamate transporter EAAT1. These results identify a new human translocon and provide a molecular framework for understanding its role in multi-pass membrane protein biogenesis. | ||||||||||||||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 6w6l.cif.gz | 5.4 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb6w6l.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 6w6l.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/w6/6w6l ftp://data.pdbj.org/pub/pdb/validation_reports/w6/6w6l | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 21435MC M: map data used to model this data C: citing same article ( |
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| Similar structure data |
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Links
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Assembly
| Deposited unit | ![]()
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Components
+60S ribosomal protein ... , 41 types, 41 molecules ABCFGHIJKLMNOPQRSTUVWXYZabcdef...
-RNA chain , 5 types, 5 molecules DEtuv
| #4: RNA chain | Mass: 50449.812 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: GenBank: 555853 |
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| #5: RNA chain | Mass: 38385.750 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) |
| #44: RNA chain | Mass: 1159525.750 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) |
| #45: RNA chain | Mass: 24414.451 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) |
| #46: RNA chain | Mass: 24436.508 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) |
-Protein/peptide , 1 types, 1 molecules y
| #47: Protein/peptide | Mass: 2315.846 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) |
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-Protein transport protein Sec61 subunit ... , 3 types, 3 molecules 123
| #48: Protein | Mass: 52295.379 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P61619 |
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| #49: Protein | Mass: 7752.325 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P60059 |
| #50: Protein | Mass: 9987.456 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: P60468 |
-Protein , 4 types, 4 molecules 4576
| #51: Protein | Mass: 25279.848 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9BVK8 |
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| #52: Protein | Mass: 63047.145 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q969V3 |
| #53: Protein | Mass: 55946.762 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q96A33 |
| #54: Protein | Mass: 21205.008 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Homo sapiens (human) / References: UniProt: Q9UM00 |
-Non-polymers , 3 types, 32 molecules 




| #55: Chemical | ChemComp-MG / #56: Chemical | #57: Water | ChemComp-HOH / | |
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-Details
| Has ligand of interest | N |
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| Has protein modification | Y |
-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 ribosome-TMCO1 translocon / Type: COMPLEX / Entity ID: #1-#54 / Source: NATURAL | |||||||||||||||||||||||||
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| Molecular weight | Value: 4 MDa / Experimental value: NO | |||||||||||||||||||||||||
| Source (natural) | Organism: Homo sapiens (human) | |||||||||||||||||||||||||
| Buffer solution | pH: 7.4 | |||||||||||||||||||||||||
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| Specimen | Conc.: 0.4 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: The sample was well dispersed on a thin carbon film | |||||||||||||||||||||||||
| Specimen support | Grid material: COPPER / Grid mesh size: 200 divisions/in. / Grid type: C-flat-1.2/1.3 | |||||||||||||||||||||||||
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 298 K Details: Two filter papers were added to each arm, 2.5 microliters of sample were added to the grids, and 0.5 seconds of drain time was allowed before vitrification. |
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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 |
| Image recording | Average exposure time: 3.8 sec. / Electron dose: 50 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of grids imaged: 1 / Num. of real images: 5562 |
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Processing
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 1049128 | ||||||||||||||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.84 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 82684 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||
| Atomic model building | Space: REAL Details: Starting models: 60S ribosomal subunit, tRNAs and nascent chain from 6om0. Sec61 heterotrimer from 6fti. Nicalin, TMEM147 are iTasser homology models based on 5a63. TMCO1 and CCDC47 are co- ...Details: Starting models: 60S ribosomal subunit, tRNAs and nascent chain from 6om0. Sec61 heterotrimer from 6fti. Nicalin, TMEM147 are iTasser homology models based on 5a63. TMCO1 and CCDC47 are co-evolution-based models from RaptorX-Contact. | ||||||||||||||||||||||||||||||||||||||||
| Atomic model building | 3D fitting-ID: 1 / Source name: PDB / Type: experimental model
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About Yorodumi




Homo sapiens (human)
United States, 5items
Citation
UCSF Chimera










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