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Yorodumi- EMDB-22716: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polycl... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-22716 | |||||||||
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Title | nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2465 | |||||||||
Map data | nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2465 | |||||||||
Sample |
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Function / homology | Function and homology information positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / positive regulation of establishment of T cell polarity / host cell endosome membrane / clathrin-dependent endocytosis of virus by host cell / viral protein processing / fusion of virus membrane with host plasma membrane / virus-mediated perturbation of host defense response / fusion of virus membrane with host endosome membrane / viral envelope ...positive regulation of plasma membrane raft polarization / positive regulation of receptor clustering / positive regulation of establishment of T cell polarity / host cell endosome membrane / clathrin-dependent endocytosis of virus by host cell / viral protein processing / fusion of virus membrane with host plasma membrane / virus-mediated perturbation of host defense response / fusion of virus membrane with host endosome membrane / viral envelope / virion attachment to host cell / apoptotic process / host cell plasma membrane / structural molecule activity / virion membrane / identical protein binding / plasma membrane Similarity search - Function | |||||||||
Biological species | synthetic construct (others) | |||||||||
Method | single particle reconstruction / negative staining / Resolution: 15.2 Å | |||||||||
Authors | Ward AB / Antanasijevic A | |||||||||
Funding support | United States, 1 items
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Citation | Journal: NPJ Vaccines / Year: 2021 Title: Immunofocusing and enhancing autologous Tier-2 HIV-1 neutralization by displaying Env trimers on two-component protein nanoparticles. Authors: Philip J M Brouwer / Aleksandar Antanasijevic / Marlon de Gast / Joel D Allen / Tom P L Bijl / Anila Yasmeen / Rashmi Ravichandran / Judith A Burger / Gabriel Ozorowski / Jonathan L Torres / ...Authors: Philip J M Brouwer / Aleksandar Antanasijevic / Marlon de Gast / Joel D Allen / Tom P L Bijl / Anila Yasmeen / Rashmi Ravichandran / Judith A Burger / Gabriel Ozorowski / Jonathan L Torres / Celia LaBranche / David C Montefiori / Rajesh P Ringe / Marit J van Gils / John P Moore / Per Johan Klasse / Max Crispin / Neil P King / Andrew B Ward / Rogier W Sanders / Abstract: The HIV-1 envelope glycoprotein trimer is poorly immunogenic because it is covered by a dense glycan shield. As a result, recombinant Env glycoproteins generally elicit inadequate antibody levels ...The HIV-1 envelope glycoprotein trimer is poorly immunogenic because it is covered by a dense glycan shield. As a result, recombinant Env glycoproteins generally elicit inadequate antibody levels that neutralize clinically relevant, neutralization-resistant (Tier-2) HIV-1 strains. Multivalent antigen presentation on nanoparticles is an established strategy to increase vaccine-driven immune responses. However, due to nanoparticle instability in vivo, the display of non-native Env structures, and the inaccessibility of many neutralizing antibody (NAb) epitopes, the effects of nanoparticle display are generally modest for Env trimers. Here, we generate two-component self-assembling protein nanoparticles presenting twenty SOSIP trimers of the clade C Tier-2 genotype 16055. We show in a rabbit immunization study that these nanoparticles induce 60-fold higher autologous Tier-2 NAb titers than the corresponding SOSIP trimers. Epitope mapping studies reveal that the presentation of 16055 SOSIP trimers on these nanoparticle focuses antibody responses to an immunodominant apical epitope. Thus, these nanoparticles are a promising platform to improve the immunogenicity of Env trimers with apex-proximate NAb epitopes. | |||||||||
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_22716.map.gz | 33.1 MB | EMDB map data format | |
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Header (meta data) | emd-22716-v30.xml emd-22716.xml | 12.6 KB 12.6 KB | Display Display | EMDB header |
Images | emd_22716.png | 54.2 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-22716 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-22716 | HTTPS FTP |
-Validation report
Summary document | emd_22716_validation.pdf.gz | 302.1 KB | Display | EMDB validaton report |
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Full document | emd_22716_full_validation.pdf.gz | 301.7 KB | Display | |
Data in XML | emd_22716_validation.xml.gz | 6.1 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22716 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22716 | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_22716.map.gz / Format: CCP4 / Size: 42.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2465 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.77 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 : nsEM map of the 16055 SOSIP HIV Env trimer in complex with polycl...
Entire | Name: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2465 (Week 22) |
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Components |
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-Supramolecule #1: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polycl...
Supramolecule | Name: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2465 (Week 22) type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1 |
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Source (natural) | Organism: synthetic construct (others) |
Recombinant expression | Organism: Homo sapiens (human) / Recombinant strain: HEK293F / Recombinant plasmid: pPPI4 |
-Experimental details
-Structure determination
Method | negative staining |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.05 mg/mL | |||||||||
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Buffer | pH: 7.4 Component:
Details: TBS buffer, pH 7.4 | |||||||||
Staining | Type: NEGATIVE / Material: Uranyl Formate Details: Sample diluted to 0.05 mg/mL. 3 uL was applied onto the grid, blotted off, and then stained with 2% uranyl formate for 60 seconds. | |||||||||
Grid | Model: Homemade / Material: COPPER / Mesh: 400 / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE | |||||||||
Details | 16055 SOSIP.v8.3 was complexed with the purified polyclonal Fab overnight. The complex was purified using SEC and concentrated. The complex was then diluted in TBS to 0.05mg/ml to be loaded onto an imaging grid. |
-Electron microscopy
Microscope | FEI TECNAI F20 |
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Image recording | Film or detector model: TVIPS TEMCAM-F416 (4k x 4k) / Number grids imaged: 1 / Average electron dose: 25.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 1.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 62000 |
Experimental equipment | Model: Tecnai F20 / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Details | No fitting was performed |
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