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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-5371 | |||||||||
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Title | Cryo-EM structure of full-length NSF in the ADP state | |||||||||
![]() | NSF hexamer in the ADP state | |||||||||
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![]() | membrane fusion / AAA+ ATPase | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 10.0 Å | |||||||||
![]() | Chang LF / Chen S / Liu CC / Pan X / Jiang J / Bai XC / Xie X / Wang HW / Sui SF | |||||||||
![]() | ![]() Title: Structural characterization of full-length NSF and 20S particles. Authors: Lei-Fu Chang / Song Chen / Cui-Cui Liu / Xijiang Pan / Jiansen Jiang / Xiao-Chen Bai / Xin Xie / Hong-Wei Wang / Sen-Fang Sui / ![]() Abstract: The 20S particle, which is composed of the N-ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs) and the SNAP receptor (SNARE) complex, has an essential role in ...The 20S particle, which is composed of the N-ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs) and the SNAP receptor (SNARE) complex, has an essential role in intracellular vesicle fusion events. Using single-particle cryo-EM and negative stain EM, we reconstructed four related three-dimensional structures: Chinese hamster NSF hexamer in the ATPγS, ADP-AlFx and ADP states, and the 20S particle. These structures reveal a parallel arrangement between the D1 and D2 domains of the hexameric NSF and characterize the nucleotide-dependent conformational changes in NSF. The structure of the 20S particle shows that it holds the SNARE complex at two interaction interfaces around the C terminus and N-terminal half of the SNARE complex, respectively. These findings provide insight into the molecular mechanism underlying disassembly of the SNARE complex by NSF. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 6.5 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 7.3 KB 7.3 KB | Display Display | ![]() |
Images | ![]() | 123.5 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 77.1 KB | Display | ![]() |
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Full document | ![]() | 76.2 KB | Display | |
Data in XML | ![]() | 493 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | NSF hexamer in the ADP state | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.8 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 : N-ethylmaleimide-sensitive factor (NSF)
Entire | Name: N-ethylmaleimide-sensitive factor (NSF) |
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Components |
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-Supramolecule #1000: N-ethylmaleimide-sensitive factor (NSF)
Supramolecule | Name: N-ethylmaleimide-sensitive factor (NSF) / type: sample / ID: 1000 / Details: The sample was freshly purified. / Oligomeric state: homohexamer / Number unique components: 1 |
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-Macromolecule #1: N-ethylmaleimide-sensitive factor
Macromolecule | Name: N-ethylmaleimide-sensitive factor / type: protein_or_peptide / ID: 1 / Name.synonym: NSF / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Vitrification | Cryogen name: ETHANE / Instrument: OTHER |
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Electron microscopy
Microscope | FEI TECNAI F20 |
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Date | Nov 15, 2010 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder: CT3500 / Specimen holder model: OTHER |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |
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Image processing
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 10.0 Å / Resolution method: FSC 0.5 CUT-OFF |
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-Atomic model buiding 1
Initial model | PDB ID: |
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Refinement | Space: REAL |