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Yorodumi- EMDB-21116: Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35... -
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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-21116 | |||||||||
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| Title | Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35 dimer) | |||||||||
Map data | Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35 dimer) | |||||||||
Sample |
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| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 6.9 Å | |||||||||
Authors | Kendall AK / Jackson LP | |||||||||
| Funding support | 1 items
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Citation | Journal: Structure / Year: 2020Title: Mammalian Retromer Is an Adaptable Scaffold for Cargo Sorting from Endosomes. Authors: Amy K Kendall / Boyang Xie / Peng Xu / Jue Wang / Rodger Burcham / Meredith N Frazier / Elad Binshtein / Hui Wei / Todd R Graham / Terunaga Nakagawa / Lauren P Jackson / ![]() Abstract: Metazoan retromer (VPS26/VPS35/VPS29) associates with sorting nexins on endosomal tubules to sort proteins to the trans-Golgi network or plasma membrane. Mechanisms of metazoan retromer assembly ...Metazoan retromer (VPS26/VPS35/VPS29) associates with sorting nexins on endosomal tubules to sort proteins to the trans-Golgi network or plasma membrane. Mechanisms of metazoan retromer assembly remain undefined. We combine single-particle cryoelectron microscopy with biophysical methods to uncover multiple oligomer structures. 2D class averages reveal mammalian heterotrimers; dimers of trimers; tetramers of trimers; and flat chains. These species are further supported by biophysical solution studies. We provide reconstructions of all species, including key sub-structures (∼5 Å resolution). Local resolution variation suggests that heterotrimers and dimers adopt multiple conformations. Our structures identify a flexible, highly conserved electrostatic dimeric interface formed by VPS35 subunits. We generate structure-based mutants to disrupt this interface in vitro. Equivalent mutations in yeast demonstrate a mild cargo-sorting defect. Our data suggest the metazoan retromer is an adaptable and plastic scaffold that accommodates interactions with different sorting nexins to sort multiple cargoes from endosomes their final destinations. | |||||||||
| History |
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_21116.map.gz | 11.1 MB | EMDB map data format | |
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| Header (meta data) | emd-21116-v30.xml emd-21116.xml | 8.5 KB 8.5 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_21116_fsc.xml | 12.6 KB | Display | FSC data file |
| Images | emd_21116.png | 153.6 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21116 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21116 | HTTPS FTP |
-Validation report
| Summary document | emd_21116_validation.pdf.gz | 78.2 KB | Display | EMDB validaton report |
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| Full document | emd_21116_full_validation.pdf.gz | 77.3 KB | Display | |
| Data in XML | emd_21116_validation.xml.gz | 494 B | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21116 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21116 | HTTPS FTP |
-Related structure data
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_21116.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35 dimer) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.096 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35...
| Entire | Name: Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35 dimer) |
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| Components |
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-Supramolecule #1: Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35...
| Supramolecule | Name: Mouse retromer (VPS26/VPS35/VPS29) chain interface I (VPS35/VPS35 dimer) type: complex / ID: 1 / Parent: 0 |
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| Source (natural) | Organism: ![]() |
| Recombinant expression | Organism: ![]() |
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 0.1 mg/mL |
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| Buffer | pH: 8.2 |
| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 71.0 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 |
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