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Yorodumi- EMDB-13576: Alpha-synuclein amyloid fibres incubated with DNAJB1, Hsc70, Hsp1... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-13576 | |||||||||
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Title | Alpha-synuclein amyloid fibres incubated with DNAJB1, Hsc70, Hsp110 and ATP | |||||||||
Map data | Alpha-synuclein amyloid fibres incubated with Hsc70, DNAJB1, Apg2 and ATP show extended stretches of densely clustered chaperones. | |||||||||
Sample |
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Biological species | Homo sapiens (human) | |||||||||
Method | electron tomography / cryo EM | |||||||||
Authors | Monistrol J / Saibil HR | |||||||||
Funding support | United Kingdom, 2 items
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Citation | Journal: EMBO J / Year: 2022 Title: Cooperative amyloid fibre binding and disassembly by the Hsp70 disaggregase. Authors: Joseph George Beton / Jim Monistrol / Anne Wentink / Erin C Johnston / Anthony John Roberts / Bernd Gerhard Bukau / Bart W Hoogenboom / Helen R Saibil / Abstract: Although amyloid fibres are highly stable protein aggregates, a specific combination of human Hsp70 system chaperones can disassemble them, including fibres formed of α-synuclein, huntingtin, or Tau. ...Although amyloid fibres are highly stable protein aggregates, a specific combination of human Hsp70 system chaperones can disassemble them, including fibres formed of α-synuclein, huntingtin, or Tau. Disaggregation requires the ATPase activity of the constitutively expressed Hsp70 family member, Hsc70, together with the J domain protein DNAJB1 and the nucleotide exchange factor Apg2. Clustering of Hsc70 on the fibrils appears to be necessary for disassembly. Here we use atomic force microscopy to show that segments of in vitro assembled α-synuclein fibrils are first coated with chaperones and then undergo bursts of rapid, unidirectional disassembly. Cryo-electron tomography and total internal reflection fluorescence microscopy reveal fibrils with regions of densely bound chaperones, preferentially at one end of the fibre. Sub-stoichiometric amounts of Apg2 relative to Hsc70 dramatically increase recruitment of Hsc70 to the fibres, creating localised active zones that then undergo rapid disassembly at a rate of ~ 4 subunits per second. The observed unidirectional bursts of Hsc70 loading and unravelling may be explained by differences between the two ends of the polar fibre structure. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_13576.map.gz | 1.2 GB | EMDB map data format | |
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Header (meta data) | emd-13576-v30.xml emd-13576.xml | 11.7 KB 11.7 KB | Display Display | EMDB header |
Images | emd_13576.png | 74.7 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-13576 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13576 | HTTPS FTP |
-Validation report
Summary document | emd_13576_validation.pdf.gz | 348.7 KB | Display | EMDB validaton report |
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Full document | emd_13576_full_validation.pdf.gz | 348.2 KB | Display | |
Data in XML | emd_13576_validation.xml.gz | 5.1 KB | Display | |
Data in CIF | emd_13576_validation.cif.gz | 5.6 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13576 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-13576 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_13576.map.gz / Format: CCP4 / Size: 1.3 GB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Annotation | Alpha-synuclein amyloid fibres incubated with Hsc70, DNAJB1, Apg2 and ATP show extended stretches of densely clustered chaperones. | ||||||||||||||||||||
Voxel size | X=Y=Z: 8.52 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
-Entire : Complex of alpha-synuclein amyloid fibres, DNAJB1, Hsc70 and Hsp1...
Entire | Name: Complex of alpha-synuclein amyloid fibres, DNAJB1, Hsc70 and Hsp110/Apg2 chaperones |
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Components |
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-Supramolecule #1: Complex of alpha-synuclein amyloid fibres, DNAJB1, Hsc70 and Hsp1...
Supramolecule | Name: Complex of alpha-synuclein amyloid fibres, DNAJB1, Hsc70 and Hsp110/Apg2 chaperones type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: Homo sapiens (human) |
Recombinant expression | Organism: Escherichia coli (E. coli) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | electron tomography |
Aggregation state | filament |
-Sample preparation
Buffer | pH: 7.5 Component:
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Grid | Model: C-flat-1.2/1.3 / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: HOLEY | ||||||||||||||||||||||||
Vitrification | Cryogen name: ETHANE | ||||||||||||||||||||||||
Sectioning | Other: NO SECTIONING | ||||||||||||||||||||||||
Fiducial marker | Manufacturer: EMS / Diameter: 10 nm |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 150.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Calibrated magnification: 42000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 4.5 µm / Nominal defocus min: 2.0 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Software - Name: IMOD (ver. 4.9.0) / Number images used: 41 |
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