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Yorodumi- PDB-9fo3: Structure of a gp140 SpyTag-SpyCatcher mi3 nanoparticle including... -
+Open data
-Basic information
Entry | Database: PDB / ID: 9fo3 | |||||||||
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Title | Structure of a gp140 SpyTag-SpyCatcher mi3 nanoparticle including mi3 density only. | |||||||||
Components | CAP255 gp140 SpyTag-SpyCatcher mi3,2-dehydro-3-deoxyphosphogluconate aldolase/4-hydroxy-2-oxoglutarate aldolase,2-dehydro-3-deoxyphosphogluconate aldolase/4-hydroxy-2-oxoglutarate aldolase | |||||||||
Keywords | IMMUNE SYSTEM / Vaccine / HIV-1 / mi3 / Nanoparticle / VLP | |||||||||
Function / homology | KDPG/KHG aldolase / KDPG and KHG aldolase / Aldolase-type TIM barrel / lyase activity / 2-dehydro-3-deoxyphosphogluconate aldolase/4-hydroxy-2-oxoglutarate aldolase Function and homology information | |||||||||
Biological species | Thermotoga maritima (bacteria) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 5.3 Å | |||||||||
Authors | Woodward, J.D. / Malebo, K. / Chapman, R. | |||||||||
Funding support | South Africa, United States, 2items
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Citation | Journal: Vaccines (Basel) / Year: 2024 Title: Development of a Two-Component Nanoparticle Vaccine Displaying an HIV-1 Envelope Glycoprotein that Elicits Tier 2 Neutralising Antibodies. Authors: Kegomoditswe Malebo / Jeremy Woodward / Phindile Ximba / Qiniso Mkhize / Sanele Cingo / Thandeka Moyo-Gwete / Penny L Moore / Anna-Lise Williamson / Rosamund Chapman / Abstract: Despite treatment and other interventions, an effective prophylactic HIV vaccine is still an essential goal in the control of HIV. Inducing robust and long-lasting antibody responses is one of the ...Despite treatment and other interventions, an effective prophylactic HIV vaccine is still an essential goal in the control of HIV. Inducing robust and long-lasting antibody responses is one of the main targets of an HIV vaccine. The delivery of HIV envelope glycoproteins (Env) using nanoparticle (NP) platforms has been shown to elicit better immunogenicity than soluble HIV Env. In this paper, we describe the development of a nanoparticle-based vaccine decorated with HIV Env using the SpyCatcher/SpyTag system. The Env utilised in this study, CAP255, was derived from a transmitted founder virus isolated from a patient who developed broadly neutralising antibodies. Negative stain and cryo-electron microscopy analyses confirmed the assembly and stability of the mi3 into uniform icosahedral NPs surrounded by regularly spaced CAP255 gp140 Env trimers. A three-dimensional reconstruction of CAP255 gp140 SpyTag-SpyCatcher mi3 clearly showed Env trimers projecting from the centre of each of the pentagonal dodecahedral faces of the NP. To our knowledge, this is the first study to report the formation of SpyCatcher pentamers on the dodecahedral faces of mi3 NPs. To investigate the immunogenicity, rabbits were primed with two doses of DNA vaccines expressing the CAP255 gp150 and a mosaic subtype C Gag and boosted with three doses of the NP-developed autologous Tier 2 CAP255 neutralising antibodies (Nabs) and low levels of heterologous CAP256SU NAbs. | |||||||||
History |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Download
PDBx/mmCIF format | 9fo3.cif.gz | 2.1 MB | Display | PDBx/mmCIF format |
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PDB format | pdb9fo3.ent.gz | 1.8 MB | Display | PDB format |
PDBx/mmJSON format | 9fo3.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 9fo3_validation.pdf.gz | 1.4 MB | Display | wwPDB validaton report |
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Full document | 9fo3_full_validation.pdf.gz | 1.4 MB | Display | |
Data in XML | 9fo3_validation.xml.gz | 265.8 KB | Display | |
Data in CIF | 9fo3_validation.cif.gz | 367.1 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/fo/9fo3 ftp://data.pdbj.org/pub/pdb/validation_reports/fo/9fo3 | HTTPS FTP |
-Related structure data
Related structure data | 50615MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
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-Components
#1: Protein | Mass: 33894.777 Da / Num. of mol.: 60 Source method: isolated from a genetically manipulated source Details: Model just contains mi3 cage component of NP. Note that SpyCatcher and gp140 are not included in the model.,Model just contains mi3 cage component of NP. Note that SpyCatcher and gp140 are ...Details: Model just contains mi3 cage component of NP. Note that SpyCatcher and gp140 are not included in the model.,Model just contains mi3 cage component of NP. Note that SpyCatcher and gp140 are not included in the model.,Model just contains mi3 cage component of NP. Note that SpyCatcher and gp140 are not included in the model.,Model just contains mi3 cage component of NP. Note that SpyCatcher and gp140 are not included in the model. Source: (gene. exp.) Thermotoga maritima (bacteria) / Gene: TM_0066 / Production host: Escherichia coli BL21 (bacteria) References: UniProt: Q9WXS1, 2-dehydro-3-deoxy-phosphogluconate aldolase Has protein modification | Y | |
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-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
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EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: CAP255 gp140 SpyTag-SpyCatcher mi3 nanoparticle / Type: COMPLEX Details: CAP255 gp140 SpyTag was expressed in HEK293T cells and purified. SpyCatcher mi3 was expressed in Entity ID: all / Source: RECOMBINANT | |||||||||||||||
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Molecular weight | Value: 4.75 MDa / Experimental value: NO | |||||||||||||||
Source (natural) | Organism: Thermotoga maritima (bacteria) | |||||||||||||||
Source (recombinant) | Organism: Escherichia coli (E. coli) / Strain: BL21 | |||||||||||||||
Buffer solution | pH: 7.4 | |||||||||||||||
Buffer component |
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Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES | |||||||||||||||
Specimen support | Grid material: COPPER / Grid mesh size: 400 divisions/in. / Grid type: Quantifoil R2/2 | |||||||||||||||
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: TFS KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD / Nominal defocus max: 5000 nm / Nominal defocus min: 500 nm / Cs: 2.7 mm / Alignment procedure: COMA FREE |
Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
Image recording | Electron dose: 45 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) |
Image scans | Width: 5760 / Height: 4092 |
-Processing
EM software |
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CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||
Symmetry | Point symmetry: I (icosahedral) | ||||||||||||||||||||
3D reconstruction | Resolution: 5.3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 1439 / Symmetry type: POINT | ||||||||||||||||||||
Atomic model building | Protocol: AB INITIO MODEL / Space: REAL / Target criteria: Cross-correlation coefficient | ||||||||||||||||||||
Atomic model building | Source name: AlphaFold / Type: in silico model |