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Yorodumi- EMDB-21435: Single Particle Cryo-EM Structure of the Natively Isolated Sec61,... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-21435 | |||||||||
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Title | Single Particle Cryo-EM Structure of the Natively Isolated Sec61, TMCO1, Nicalin, TMEM147, and CCDC47 Containing Ribosome-Translocon Complex After Focused Refinement with Local Angular Searches | |||||||||
Map data | ||||||||||
Sample |
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Function / homology | Function and homology information negative regulation of nodal signaling pathway / multi-pass transmembrane protein insertion into ER membrane / multi-pass translocon complex / protein localization to nuclear inner membrane / endoplasmic reticulum Sec complex / determination of left/right asymmetry in lateral mesoderm / pronephric nephron development / rough endoplasmic reticulum membrane / endoplasmic reticulum quality control compartment / cotranslational protein targeting to membrane ...negative regulation of nodal signaling pathway / multi-pass transmembrane protein insertion into ER membrane / multi-pass translocon complex / protein localization to nuclear inner membrane / endoplasmic reticulum Sec complex / determination of left/right asymmetry in lateral mesoderm / pronephric nephron development / rough endoplasmic reticulum membrane / endoplasmic reticulum quality control compartment / cotranslational protein targeting to membrane / Ssh1 translocon complex / Sec61 translocon complex / protein targeting to ER / protein insertion into ER membrane / regulation of protein complex stability / post-translational protein targeting to endoplasmic reticulum membrane / SRP-dependent cotranslational protein targeting to membrane, translocation / eukaryotic 80S initiation complex / negative regulation of protein neddylation / translation at presynapse / axial mesoderm development / embryonic brain development / endoplasmic reticulum calcium ion homeostasis / negative regulation of formation of translation preinitiation complex / 90S preribosome assembly / SRP-dependent cotranslational protein targeting to membrane / signal sequence binding / post-translational protein targeting to membrane, translocation / endoplasmic reticulum organization / TORC2 complex binding / GAIT complex / middle ear morphogenesis / protein folding chaperone complex / A band / alpha-beta T cell differentiation / cytoplasmic side of rough endoplasmic reticulum membrane / retrograde protein transport, ER to cytosol / epidermal growth factor binding / exit from mitosis / positive regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator / regulation of translation involved in cellular response to UV / positive regulation of DNA damage response, signal transduction by p53 class mediator resulting in transcription of p21 class mediator / negative regulation of ubiquitin protein ligase activity / optic nerve development / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / response to aldosterone / retinal ganglion cell axon guidance / homeostatic process / ER overload response / macrophage chemotaxis / lung morphogenesis / Protein hydroxylation / Peptide chain elongation / Selenocysteine synthesis / positive regulation of signal transduction by p53 class mediator / Formation of a pool of free 40S subunits / blastocyst development / ubiquitin ligase inhibitor activity / Eukaryotic Translation Termination / Response of EIF2AK4 (GCN2) to amino acid deficiency / protein transmembrane transporter activity / SRP-dependent cotranslational protein targeting to membrane / Viral mRNA Translation / protein localization to nucleus / Nonsense Mediated Decay (NMD) independent of the Exon Junction Complex (EJC) / GTP hydrolysis and joining of the 60S ribosomal subunit / regulation of protein-containing complex assembly / regulation of signal transduction / L13a-mediated translational silencing of Ceruloplasmin expression / Major pathway of rRNA processing in the nucleolus and cytosol / protein targeting / protein-RNA complex assembly / cellular response to interleukin-4 / Nonsense Mediated Decay (NMD) enhanced by the Exon Junction Complex (EJC) / negative regulation of ubiquitin-dependent protein catabolic process / translation regulator activity / rough endoplasmic reticulum / ERAD pathway / MDM2/MDM4 family protein binding / negative regulation of proteasomal ubiquitin-dependent protein catabolic process / maturation of LSU-rRNA / protein folding chaperone / DNA damage response, signal transduction by p53 class mediator resulting in cell cycle arrest / cytosolic ribosome / embryo implantation / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / post-embryonic development / ossification / ribosomal large subunit biogenesis / regulation of signal transduction by p53 class mediator / guanyl-nucleotide exchange factor activity / innate immune response in mucosa / skeletal system development / positive regulation of translation / mRNA 3'-UTR binding / mitochondrial membrane / sensory perception of sound / calcium ion transmembrane transport / bone development / calcium channel activity Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.8 Å | |||||||||
Authors | McGilvray PT / Anghel SA / Sundaram A / Trnka MJ / Zhong F / Hu H / Burlingame AL / Keenan RJ | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Elife / Year: 2020 Title: 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. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_21435.map.gz | 412.1 MB | EMDB map data format | |
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Header (meta data) | emd-21435-v30.xml emd-21435.xml | 66.1 KB 66.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_21435_fsc.xml | 17.6 KB | Display | FSC data file |
Images | emd_21435.png | 140.8 KB | ||
Masks | emd_21435_msk_1.map | 476.8 MB | Mask map | |
Others | emd_21435_half_map_1.map.gz emd_21435_half_map_2.map.gz | 416.1 MB 416.8 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21435 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21435 | HTTPS FTP |
-Validation report
Summary document | emd_21435_validation.pdf.gz | 516.9 KB | Display | EMDB validaton report |
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Full document | emd_21435_full_validation.pdf.gz | 516.5 KB | Display | |
Data in XML | emd_21435_validation.xml.gz | 22.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21435 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21435 | HTTPS FTP |
-Related structure data
Related structure data | 6w6lMC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_21435.map.gz / Format: CCP4 / Size: 476.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.36 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_21435_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_21435_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_21435_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Native complex of a translating ribosome and a novel translocon c...
+Supramolecule #1: Native complex of a translating ribosome and a novel translocon c...
+Supramolecule #2: 60S Subunit
+Supramolecule #3: 40S Subunit
+Supramolecule #4: Translocon
+Supramolecule #5: Sec61 Complex
+Supramolecule #6: TMCO1
+Supramolecule #7: NOMO-Nicalin-TMEM147 Complex
+Supramolecule #8: CCDC47
+Supramolecule #9: 60S acidic ribosomal protein P0
+Supramolecule #10: 60S acidic ribosomal protein P1
+Supramolecule #11: 60S acidic ribosomal protein P2
+Supramolecule #12: 60S ribosomal protein L10
+Supramolecule #13: 60S ribosomal protein L10a
+Supramolecule #14: 60S ribosomal protein L11
+Supramolecule #15: 60S ribosomal protein L12
+Supramolecule #16: 60S ribosomal protein L13
+Supramolecule #17: 60S ribosomal protein L13a
+Supramolecule #18: 60S ribosomal protein L14
+Supramolecule #19: 60S ribosomal protein L15
+Supramolecule #20: 60S ribosomal protein L17
+Supramolecule #21: 60S ribosomal protein L18
+Supramolecule #22: 60S ribosomal protein L18a
+Supramolecule #23: 60S ribosomal protein L19
+Supramolecule #24: 60S ribosomal protein L21
+Supramolecule #25: 60S ribosomal protein L22
+Supramolecule #26: 60S ribosomal protein L23
+Supramolecule #27: 60S ribosomal protein L23a
+Supramolecule #28: 60S ribosomal protein L24
+Supramolecule #29: 60S ribosomal protein L26
+Supramolecule #30: 60S ribosomal protein L27
+Supramolecule #31: 60S ribosomal protein L27a
+Supramolecule #32: 60S ribosomal protein L28
+Supramolecule #33: 60S ribosomal protein L3
+Supramolecule #34: 60S ribosomal protein L30
+Supramolecule #35: 60S ribosomal protein L31
+Supramolecule #36: 60S ribosomal protein L32
+Supramolecule #37: 60S ribosomal protein L34
+Supramolecule #38: 60S ribosomal protein L35
+Supramolecule #39: 60S ribosomal protein L35a
+Supramolecule #40: 60S ribosomal protein L36
+Supramolecule #41: 60S ribosomal protein L36a
+Supramolecule #42: 60S ribosomal protein L37a
+Supramolecule #43: 60S ribosomal protein L38
+Supramolecule #44: 60S ribosomal protein L39
+Supramolecule #45: 60S ribosomal protein L4
+Supramolecule #46: 60S ribosomal protein L5
+Supramolecule #47: 60S ribosomal protein L6
+Supramolecule #48: 60S ribosomal protein L7
+Supramolecule #49: 60S ribosomal protein L7a
+Supramolecule #50: 60S ribosomal protein L8
+Supramolecule #51: 60S ribosomal protein L9
+Supramolecule #52: 40S ribosomal protein S10
+Supramolecule #53: 40S ribosomal protein S11
+Supramolecule #54: 40S ribosomal protein S12
+Supramolecule #55: 40S ribosomal protein S13
+Supramolecule #56: 40S ribosomal protein S14
+Supramolecule #57: 40S ribosomal protein S15
+Supramolecule #58: 40S ribosomal protein S15a
+Supramolecule #59: 40S ribosomal protein S16
+Supramolecule #60: 40S ribosomal protein S17
+Supramolecule #61: 40S ribosomal protein S18
+Supramolecule #62: 40S ribosomal protein S19
+Supramolecule #63: 40S ribosomal protein S2
+Supramolecule #64: 40S ribosomal protein S20
+Supramolecule #65: 40S ribosomal protein S21
+Supramolecule #66: 40S ribosomal protein S23
+Supramolecule #67: 40S ribosomal protein S24
+Supramolecule #68: 40S ribosomal protein S25
+Supramolecule #69: 40S ribosomal protein S26
+Supramolecule #70: 40S ribosomal protein S27
+Supramolecule #71: 40S ribosomal protein S27-like
+Supramolecule #72: 40S ribosomal protein S28
+Supramolecule #73: 40S ribosomal protein S29
+Supramolecule #74: 40S ribosomal protein S3
+Supramolecule #75: 40S ribosomal protein S30
+Supramolecule #76: 40S ribosomal protein S3a
+Supramolecule #77: 40S ribosomal protein S4, X isoform
+Supramolecule #78: 40S ribosomal protein S5
+Supramolecule #79: 40S ribosomal protein S6
+Supramolecule #80: 40S ribosomal protein S7
+Supramolecule #81: 40S ribosomal protein S8
+Supramolecule #82: 40S ribosomal protein S9
+Supramolecule #83: 40S ribosomal protein SA
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.4 mg/mL | |||||||||||||||
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Buffer | pH: 7.4 Component:
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Grid | Model: C-flat-1.2/1.3 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Support film - Film thickness: 2.0 nm / Pretreatment - Type: GLOW DISCHARGE / Details: unspecified | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 298 K / Instrument: FEI VITROBOT MARK IV Details: Two filter papers were added to each arm, 2.5 microliters of sample were added to grids, and 0.5 second of drain time was allowed before vitrification.. | |||||||||||||||
Details | Sample was well-dispersed on a thin (~2nm) carbon film. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | #0 - Image recording ID: 1 / #0 - Film or detector model: GATAN K3 (6k x 4k) / #0 - Detector mode: COUNTING / #0 - Number grids imaged: 1 / #0 - Number real images: 5562 / #0 - Average exposure time: 3.8 sec. / #0 - Average electron dose: 50.0 e/Å2 / #1 - Image recording ID: 2 / #1 - Film or detector model: GATAN K3 (6k x 4k) / #1 - Number grids imaged: 1 / #1 - Number real images: 5562 / #1 - Average exposure time: 3.8 sec. / #1 - Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |