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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-2874 | |||||||||
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Title | Cryo electron microscopy of SNAP-SNARE assembly in 20S particle | |||||||||
![]() | Reconstruction of alpha-SNAP-SNARE assembly in 20S particle | |||||||||
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![]() | 20S particles / SNARE / alpha-SNAP / membrane fusion | |||||||||
Biological species | ![]() ![]() ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 7.35 Å | |||||||||
![]() | Zhou Q / Huang X / Sun S / Li XM / Wang HW / Sui SF | |||||||||
![]() | ![]() Title: Cryo-EM structure of SNAP-SNARE assembly in 20S particle. Authors: Qiang Zhou / Xuan Huang / Shan Sun / Xueming Li / Hong-Wei Wang / Sen-Fang Sui / ![]() Abstract: N-ethylmaleimide-sensitive factor (NSF) and α soluble NSF attachment proteins (α-SNAPs) work together within a 20S particle to disassemble and recycle the SNAP receptor (SNARE) complex after ...N-ethylmaleimide-sensitive factor (NSF) and α soluble NSF attachment proteins (α-SNAPs) work together within a 20S particle to disassemble and recycle the SNAP receptor (SNARE) complex after intracellular membrane fusion. To understand the disassembly mechanism of the SNARE complex by NSF and α-SNAP, we performed single-particle cryo-electron microscopy analysis of 20S particles and determined the structure of the α-SNAP-SNARE assembly portion at a resolution of 7.35 Å. The structure illustrates that four α-SNAPs wrap around the single left-handed SNARE helical bundle as a right-handed cylindrical assembly within a 20S particle. A conserved hydrophobic patch connecting helices 9 and 10 of each α-SNAP forms a chock protruding into the groove of the SNARE four-helix bundle. Biochemical studies proved that this structural element was critical for SNARE complex disassembly. Our study suggests how four α-SNAPs may coordinate with the NSF to tear the SNARE complex into individual proteins. | |||||||||
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 | ![]() | 7.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 12.4 KB 12.4 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 4.5 KB | Display | ![]() |
Images | ![]() | 89.9 KB | ||
Masks | ![]() | 8 MB | ![]() | |
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 274 KB | Display | ![]() |
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Full document | ![]() | 273.1 KB | Display | |
Data in XML | ![]() | 7.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Reconstruction of alpha-SNAP-SNARE assembly in 20S particle | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.32 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Segmentation: This is a soft spherical mask.
Annotation | This is a soft spherical mask. | ||||||||||||
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File | ![]() | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : alpha-SNAP-SNARE assembly in 20S particle
Entire | Name: alpha-SNAP-SNARE assembly in 20S particle |
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Components |
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-Supramolecule #1000: alpha-SNAP-SNARE assembly in 20S particle
Supramolecule | Name: alpha-SNAP-SNARE assembly in 20S particle / type: sample / ID: 1000 / Details: 20S particle was prepared in nanodisc. Oligomeric state: One homotetramer of alpha-SNAP binds to one monomer of SNARE complex. Number unique components: 2 |
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Molecular weight | Theoretical: 200 KDa |
-Macromolecule #1: soluble NSF attachment protein receptor
Macromolecule | Name: soluble NSF attachment protein receptor / type: protein_or_peptide / ID: 1 / Name.synonym: SNARE / Number of copies: 1 / Oligomeric state: monomer / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 66 KDa |
Recombinant expression | Organism: ![]() ![]() |
-Macromolecule #2: alpha soluble NSF attachment protein
Macromolecule | Name: alpha soluble NSF attachment protein / type: protein_or_peptide / ID: 2 / Name.synonym: alpha-SNAP / Number of copies: 4 / Oligomeric state: tetramer / Recombinant expression: Yes |
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Source (natural) | Organism: ![]() ![]() |
Molecular weight | Theoretical: 33 KDa |
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
Concentration | 0.2 mg/mL |
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Grid | Details: 400 mesh copper grid with thin carbon support |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Instrument: FEI VITROBOT MARK IV / Method: Blot for 1 second before plunging |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Alignment procedure | Legacy - Astigmatism: Objective lens astigmatism was corrected at 22,500 times magnification. |
Date | Jul 2, 2014 |
Image recording | Category: CCD / Film or detector model: GATAN K2 (4k x 4k) / Number real images: 2349 / Average electron dose: 46 e/Å2 Details: Every image is the average of motion-corrected movie frames. |
Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 22500 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - #0 - Chain ID: A / Chain - #1 - Chain ID: B / Chain - #2 - Chain ID: C / Chain - #3 - Chain ID: D |
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Software | Name: ![]() |
Refinement | Space: REAL / Protocol: RIGID BODY FIT |