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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 | |||||||||
Map data | NSF hexamer in the ADP state | |||||||||
Sample |
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Keywords | membrane fusion / AAA+ ATPase | |||||||||
| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 10.0 Å | |||||||||
Authors | Chang LF / Chen S / Liu CC / Pan X / Jiang J / Bai XC / Xie X / Wang HW / Sui SF | |||||||||
Citation | Journal: Nat Struct Mol Biol / Year: 2012Title: 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 |
Movie viewer |
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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_5371.map.gz | 6.5 MB | EMDB map data format | |
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| Header (meta data) | emd-5371-v30.xml emd-5371.xml | 7.3 KB 7.3 KB | Display Display | EMDB header |
| Images | emd_5371_1.jpg | 123.5 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-5371 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-5371 | HTTPS FTP |
-Validation report
| Summary document | emd_5371_validation.pdf.gz | 77.1 KB | Display | EMDB validaton report |
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| Full document | emd_5371_full_validation.pdf.gz | 76.2 KB | Display | |
| Data in XML | emd_5371_validation.xml.gz | 493 B | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5371 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5371 | 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_5371.map.gz / Format: CCP4 / Size: 7.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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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Processing | 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: FIELD EMISSION GUN |
| 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 |
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