+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-23737 | |||||||||
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Title | PolyQ40 oligomer tomogram at 2uM concentration | |||||||||
Map data | PolyQ40 oligomer | |||||||||
Sample |
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Biological species | Homo sapiens (human) | |||||||||
Method | electron tomography / cryo EM | |||||||||
Authors | Nunn K / Jiang J / Dai W | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Biochem Biophys Res Commun / Year: 2021 Title: Structural insight into transmissive mutant huntingtin species by correlative light and electron microscopy and cryo-electron tomography. Authors: Xuyuan Kuang / Kyle Nunn / Jennifer Jiang / Paul Castellano / Uttara Hardikar / Arianna Horgan / Joyce Kong / Zhiqun Tan / Wei Dai / Abstract: Aggregates of mutant huntingtin (mHTT) containing an expanded polyglutamine (polyQ) tract are hallmarks of Huntington's Disease (HD). Studies have shown that mHTT can spread between cells, leading to ...Aggregates of mutant huntingtin (mHTT) containing an expanded polyglutamine (polyQ) tract are hallmarks of Huntington's Disease (HD). Studies have shown that mHTT can spread between cells, leading to the propagation of misfolded protein pathology. However, the structure of transmissive mHTT species, and the molecular mechanisms underlying their transmission remain unknown. Using correlative light and electron microscopy (CLEM) and cryo-electron tomography (cryo-ET), we identified two types of aggregation-prone granules in conditioned medium from PC12 cells expressing a mHTT N-terminal fragment: densities enclosed by extracellular vesicles (EVs), and uncoated, amorphous meshworks of heterogeneous oligomers that co-localize with clusters of EVs. In vitro assays confirmed that liposomes induce condensation of polyQ oligomers into higher-order assemblies, resembling the uncoated meshworks observed in PC12 conditioned medium. Our findings provide novel insights into formation and architecture of transmissive mHTT proteins, and highlight the potential role of EVs as both carriers and modulators of transmissive mHTT proteins. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_23737.map.gz | 46.8 MB | EMDB map data format | |
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Header (meta data) | emd-23737-v30.xml emd-23737.xml | 10 KB 10 KB | Display Display | EMDB header |
Images | emd_23737.png | 47.7 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-23737 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-23737 | HTTPS FTP |
-Validation report
Summary document | emd_23737_validation.pdf.gz | 255.1 KB | Display | EMDB validaton report |
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Full document | emd_23737_full_validation.pdf.gz | 254.7 KB | Display | |
Data in XML | emd_23737_validation.xml.gz | 4.7 KB | Display | |
Data in CIF | emd_23737_validation.cif.gz | 5.2 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23737 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-23737 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_23737.map.gz / Format: CCP4 / Size: 50.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||
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Annotation | PolyQ40 oligomer | ||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. generated in cubic-lattice coordinate | ||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 6.986 Å | ||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
-Entire : Synthetic polyQ40 oligomers
Entire | Name: Synthetic polyQ40 oligomers |
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Components |
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-Supramolecule #1: Synthetic polyQ40 oligomers
Supramolecule | Name: Synthetic polyQ40 oligomers / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Homo sapiens (human) |
-Macromolecule #1: Synthetic polyQ peptide
Macromolecule | Name: Synthetic polyQ peptide / type: other / ID: 1 / Classification: other |
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Source (natural) | Organism: Homo sapiens (human) |
Sequence | String: KKQQQQQQQQ QQQQQQQQQQ QQQQQQQQQQ QQQQQQQQQQ QQKK |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | electron tomography |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7 |
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 293.15 K / Instrument: LEICA EM GP |
Sectioning | Other: NO SECTIONING |
Fiducial marker | Manufacturer: EMS / Diameter: 10 nm |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 1.5 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal magnification: 31000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Software - Name: IMOD / Number images used: 47 |
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