+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9725 | |||||||||
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Title | state3 of the whole 20S complex | |||||||||
Map data | ||||||||||
Sample |
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Biological species | Cricetulus griseus (Chinese hamster) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 8.241905 Å | |||||||||
Authors | Huang X / Wang X / Sun S / Sui SF | |||||||||
Citation | Journal: Sci Adv / Year: 2019 Title: Mechanistic insights into the SNARE complex disassembly. Authors: Xuan Huang / Shan Sun / Xiaojing Wang / Fenghui Fan / Qiang Zhou / Shan Lu / Yong Cao / Qiu-Wen Wang / Meng-Qiu Dong / Jun Yao / Sen-Fang Sui / Abstract: NSF (-ethylmaleimide-sensitive factor) and α-SNAP (α-soluble NSF attachment protein) bind to the SNARE (soluble NSF attachment protein receptor) complex, the minimum machinery to mediate membrane ...NSF (-ethylmaleimide-sensitive factor) and α-SNAP (α-soluble NSF attachment protein) bind to the SNARE (soluble NSF attachment protein receptor) complex, the minimum machinery to mediate membrane fusion, to form a 20S complex, which disassembles the SNARE complex for reuse. We report the cryo-EM structures of the α-SNAP-SNARE subcomplex and the NSF-D1D2 domain in the 20S complex at 3.9- and 3.7-Å resolutions, respectively. Combined with the biochemical and electrophysiological analyses, we find that α-SNAPs use R116 through electrostatic interactions and L197 through hydrophobic interactions to apply force mainly on two positions of the VAMP protein to execute disassembly process. Furthermore, we define the interaction between the amino terminus of the SNARE helical bundle and the pore loop of the NSF-D1 domain and demonstrate its essential role as a potential anchor for SNARE complex disassembly. Our studies provide a rotation model of α-SNAP-mediated disassembly of the SNARE complex. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_9725.map.gz | 49.6 MB | EMDB map data format | |
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Header (meta data) | emd-9725-v30.xml emd-9725.xml | 8.8 KB 8.8 KB | Display Display | EMDB header |
Images | emd_9725.png | 122 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9725 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9725 | HTTPS FTP |
-Validation report
Summary document | emd_9725_validation.pdf.gz | 78.6 KB | Display | EMDB validaton report |
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Full document | emd_9725_full_validation.pdf.gz | 77.7 KB | Display | |
Data in XML | emd_9725_validation.xml.gz | 495 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9725 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9725 | HTTPS FTP |
-Related structure data
Related structure data | 9697C 9698C 9723C 9724C 9726C 9727C 9728C 9729C 6ip1C 6ip2C C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_9725.map.gz / Format: CCP4 / Size: 64 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.30654 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : state3 of the whole 20S complex
Entire | Name: state3 of the whole 20S complex |
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Components |
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-Supramolecule #1: state3 of the whole 20S complex
Supramolecule | Name: state3 of the whole 20S complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: Cricetulus griseus (Chinese hamster) |
Recombinant expression | Organism: Escherichia coli (E. coli) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 8.241905 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 17668 |
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Initial angle assignment | Type: RANDOM ASSIGNMENT |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |