+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-5370 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-EM structure of full-length NSF in the ADP-AlFx state | |||||||||
Map data | NSF hexamer in the ADP-AlFx state | |||||||||
Sample |
| |||||||||
Keywords | membrane fusion / AAA+ ATPase | |||||||||
Biological species | Cricetulus griseus (Chinese hamster) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 9.2 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: 2012 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 |
|
-Structure visualization
Movie |
Movie viewer |
---|---|
Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_5370.map.gz | 6.5 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-5370-v30.xml emd-5370.xml | 7.3 KB 7.3 KB | Display Display | EMDB header |
Images | emd_5370_1.jpg | 181.4 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-5370 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-5370 | HTTPS FTP |
-Validation report
Summary document | emd_5370_validation.pdf.gz | 77.4 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_5370_full_validation.pdf.gz | 76.5 KB | Display | |
Data in XML | emd_5370_validation.xml.gz | 492 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5370 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-5370 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|
-Map
File | Download / File: emd_5370.map.gz / Format: CCP4 / Size: 7.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | NSF hexamer in the ADP-AlFx state | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.8 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-Sample components
-Entire : N-ethylmaleimide-sensitive factor (NSF)
Entire | Name: N-ethylmaleimide-sensitive factor (NSF) |
---|---|
Components |
|
-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 |
---|
-Macromolecule #1: N-ethylmaleimide-sensitive factor
Macromolecule | Name: N-ethylmaleimide-sensitive factor / type: protein_or_peptide / ID: 1 / Name.synonym: NSF / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: Cricetulus griseus (Chinese hamster) / synonym: Chinese hamster |
Recombinant expression | Organism: Escherichia coli (E. coli) |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Vitrification | Cryogen name: ETHANE / Instrument: OTHER |
---|
-Electron microscopy
Microscope | FEI TECNAI F20 |
---|---|
Date | Sep 29, 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 |
-Image processing
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 9.2 Å / Resolution method: FSC 0.5 CUT-OFF |
---|
-Atomic model buiding 1
Initial model | PDB ID: |
---|---|
Refinement | Space: REAL |