[English] 日本語
Yorodumi- EMDB-3072: Representative tomogram of canine pancreatic rough Endoplasmic Re... -
+
Open data
-
Basic information
| Entry | Database: EMDB / ID: EMD-3072 | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Title | Representative tomogram of canine pancreatic rough Endoplasmic Reticulum vesicles | |||||||||
Map data | Representative tomogram of canine pancreatic rough Endoplasmic Reticulum vesicles. | |||||||||
Sample |
| |||||||||
Keywords | Ribosome / Sec61 / Translocon / Endoplasmic Reticulum / Cryoelectron Tomography / Subtomogram Analysis | |||||||||
| Biological species | ![]() | |||||||||
| Method | electron tomography / cryo EM | |||||||||
Authors | Pfeffer S / Burbaum L / Unverdorben P / Pech M / Chen Y / Zimmermann R / Beckmann R / Foerster F | |||||||||
Citation | Journal: Nat Commun / Year: 2015Title: Structure of the native Sec61 protein-conducting channel. Authors: Stefan Pfeffer / Laura Burbaum / Pia Unverdorben / Markus Pech / Yuxiang Chen / Richard Zimmermann / Roland Beckmann / Friedrich Förster / ![]() Abstract: In mammalian cells, secretory and membrane proteins are translocated across or inserted into the endoplasmic reticulum (ER) membrane by the universally conserved protein-conducting channel Sec61, ...In mammalian cells, secretory and membrane proteins are translocated across or inserted into the endoplasmic reticulum (ER) membrane by the universally conserved protein-conducting channel Sec61, which has been structurally studied in isolated, detergent-solubilized states. Here we structurally and functionally characterize native, non-solubilized ribosome-Sec61 complexes on rough ER vesicles using cryo-electron tomography and ribosome profiling. Surprisingly, the 9-Å resolution subtomogram average reveals Sec61 in a laterally open conformation, even though the channel is not in the process of inserting membrane proteins into the lipid bilayer. In contrast to recent mechanistic models for polypeptide translocation and insertion, our results indicate that the laterally open conformation of Sec61 is the only conformation present in the ribosome-bound translocon complex, independent of its functional state. Consistent with earlier functional studies, our structure suggests that the ribosome alone, even without a nascent chain, is sufficient for lateral opening of Sec61 in a lipid environment. | |||||||||
| History |
|
-
Structure visualization
| Movie |
Movie viewer |
|---|---|
| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
-
Downloads & links
-EMDB archive
| Map data | emd_3072.map.gz | 96.6 MB | EMDB map data format | |
|---|---|---|---|---|
| Header (meta data) | emd-3072-v30.xml emd-3072.xml | 9.3 KB 9.3 KB | Display Display | EMDB header |
| Images | EMD-3072.tif | 398.5 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-3072 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3072 | HTTPS FTP |
-Validation report
| Summary document | emd_3072_validation.pdf.gz | 155.3 KB | Display | EMDB validaton report |
|---|---|---|---|---|
| Full document | emd_3072_full_validation.pdf.gz | 154.4 KB | Display | |
| Data in XML | emd_3072_validation.xml.gz | 4.6 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3072 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-3072 | HTTPS FTP |
-Related structure data
-
Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
|---|---|
| Related items in Molecule of the Month |
-
Map
| File | Download / File: emd_3072.map.gz / Format: CCP4 / Size: 101.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Annotation | Representative tomogram of canine pancreatic rough Endoplasmic Reticulum vesicles. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 20.96 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
-Supplemental data
-
Sample components
-Entire : Representative tomogram of canine pancreatic rough Endoplasmic Re...
| Entire | Name: Representative tomogram of canine pancreatic rough Endoplasmic Reticulum vesicles. |
|---|---|
| Components |
|
-Supramolecule #1000: Representative tomogram of canine pancreatic rough Endoplasmic Re...
| Supramolecule | Name: Representative tomogram of canine pancreatic rough Endoplasmic Reticulum vesicles. type: sample / ID: 1000 / Number unique components: 2 |
|---|
-Supramolecule #1: Membrane-bound 80S ribosome
| Supramolecule | Name: Membrane-bound 80S ribosome / type: complex / ID: 1 / Recombinant expression: No / Database: NCBI / Ribosome-details: ribosome-eukaryote: ALL |
|---|---|
| Source (natural) | Organism: ![]() |
-Macromolecule #1: ER protein translocon
| Macromolecule | Name: ER protein translocon / type: protein_or_peptide / ID: 1 / Recombinant expression: No / Database: NCBI |
|---|---|
| Source (natural) | Organism: ![]() |
-Experimental details
-Structure determination
| Method | cryo EM |
|---|---|
Processing | electron tomography |
| Aggregation state | particle |
-
Sample preparation
| Concentration | 2 mg/mL |
|---|---|
| Buffer | pH: 7.6 / Details: 20mM Hepes, 50mM KCl; 2mM MgCl2 |
| Grid | Details: Lacey carbon molybdenum grid |
| Vitrification | Cryogen name: ETHANE-PROPANE MIXTURE / Chamber humidity: 70 % / Instrument: FEI VITROBOT MARK IV / Method: Blot 3 seconds before plunging. |
-
Electron microscopy
| Microscope | FEI TITAN KRIOS |
|---|---|
| Specialist optics | Energy filter - Name: Gatan |
| Date | Jun 18, 2014 |
| Image recording | Category: CCD / Film or detector model: GATAN K2 (4k x 4k) / Average electron dose: 100 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 3.0 µm |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Tilt series - Axis1 - Min angle: -60 ° / Tilt series - Axis1 - Max angle: 60 ° / Tilt series - Axis1 - Angle increment: 2 ° |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
-
Image processing
| Details | Tomogram reconstruction was accomplished using PyTom. |
|---|---|
| Final reconstruction | Software - Name: PyTom, tom_toolbox, av3_toolbox / Number images used: 61 |
| CTF correction | Details: Each tilt image |
Movie
Controller
About Yorodumi


Keywords
Authors
Citation
UCSF Chimera





Z (Sec.)
Y (Row.)
X (Col.)


















