+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-8089 | |||||||||
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Title | G1S VLP_VPP | |||||||||
Map data | None | |||||||||
Sample |
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Biological species | Influenza A virus | |||||||||
Method | electron tomography / cryo EM | |||||||||
Authors | Chlanda P / Zimmerberg J | |||||||||
Citation | Journal: Nat Microbiol / Year: 2016 Title: The hemifusion structure induced by influenza virus haemagglutinin is determined by physical properties of the target membranes. Authors: Petr Chlanda / Elena Mekhedov / Hang Waters / Cindi L Schwartz / Elizabeth R Fischer / Rolf J Ryham / Fredric S Cohen / Paul S Blank / Joshua Zimmerberg / Abstract: Influenza A virus haemagglutinin conformational change drives the membrane fusion of viral and endosomal membranes at low pH. Membrane fusion proceeds through an intermediate called hemifusion(1,2). ...Influenza A virus haemagglutinin conformational change drives the membrane fusion of viral and endosomal membranes at low pH. Membrane fusion proceeds through an intermediate called hemifusion(1,2). For viral fusion, the hemifusion structures are not determined(3). Here, influenza virus-like particles(4) carrying wild-type haemagglutinin or haemagglutinin hemifusion mutant G1S(5) and liposome mixtures were studied at low pH by Volta phase plate cryo-electron tomography, which improves the signal-to-noise ratio close to focus. We determined two distinct hemifusion structures: a hemifusion diaphragm and a novel structure termed a 'lipidic junction'. Liposomes with lipidic junctions were ruptured with membrane edges stabilized by haemagglutinin. The rupture frequency and hemifusion diaphragm diameter were not affected by G1S mutation, but decreased when the cholesterol level in the liposomes was close to physiological concentrations. We propose that haemagglutinin induces a merger between the viral and target membranes by one of two independent pathways: a rupture-insertion pathway leading to the lipidic junction and a hemifusion-stalk pathway leading to a fusion pore. The latter is relevant under the conditions of influenza virus infection of cells. Cholesterol concentration functions as a pathway switch because of its negative spontaneous curvature in the target bilayer, as determined by continuum analysis. | |||||||||
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_8089.map.gz | 18.8 MB | EMDB map data format | |
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Header (meta data) | emd-8089-v30.xml emd-8089.xml | 7.9 KB 7.9 KB | Display Display | EMDB header |
Images | emd_8089.png | 238.3 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-8089 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-8089 | HTTPS FTP |
-Validation report
Summary document | emd_8089_validation.pdf.gz | 78.2 KB | Display | EMDB validaton report |
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Full document | emd_8089_full_validation.pdf.gz | 77.3 KB | Display | |
Data in XML | emd_8089_validation.xml.gz | 498 B | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8089 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-8089 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_8089.map.gz / Format: CCP4 / Size: 27 MB / Type: IMAGE STORED AS SIGNED BYTE | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | None | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 7.36 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 : Influenza A virus
Entire | Name: Influenza A virus |
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Components |
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-Supramolecule #1: Influenza A virus
Supramolecule | Name: Influenza A virus / type: virus / ID: 1 / Parent: 0 / Macromolecule list: #1 / NCBI-ID: 11320 / Sci species name: Influenza A virus / Virus type: VIRUS-LIKE PARTICLE / Virus isolate: OTHER / Virus enveloped: Yes / Virus empty: Yes |
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Host system | Organism: Homo sapiens (human) / Recombinant cell: HEK 293T / Recombinant plasmid: pCAGGS |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | electron tomography |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 5 |
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % |
Sectioning | Other: NO SECTIONING |
Fiducial marker | Manufacturer: EMS / Diameter: 100 nm |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: FEI FALCON I (4k x 4k) / Average electron dose: 1.3 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 | Algorithm: BACK PROJECTION / Software - Name: Imod / Number images used: 60 |
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