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Yorodumi- PDB-7nur: Structure of the Toxoplasma gondii kinase Ron13, kinase-dead mutant -
+Open data
-Basic information
Entry | Database: PDB / ID: 7nur | |||||||||
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Title | Structure of the Toxoplasma gondii kinase Ron13, kinase-dead mutant | |||||||||
Components | Protein kinase domain-containing protein | |||||||||
Keywords | TRANSFERASE / secretory pathway / kinase | |||||||||
Function / homology | Function and homology information | |||||||||
Biological species | Toxoplasma gondii (eukaryote) | |||||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.125 Å | |||||||||
Authors | Korkhov, V.M. / Mehta, V. | |||||||||
Funding support | Switzerland, 2items
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Citation | Journal: Nat Commun / Year: 2021 Title: Structural insights into an atypical secretory pathway kinase crucial for Toxoplasma gondii invasion. Authors: Gaëlle Lentini / Rouaa Ben Chaabene / Oscar Vadas / Chandra Ramakrishnan / Budhaditya Mukherjee / Ved Mehta / Matteo Lunghi / Jonas Grossmann / Bohumil Maco / Rémy Visentin / Adrian B Hehl ...Authors: Gaëlle Lentini / Rouaa Ben Chaabene / Oscar Vadas / Chandra Ramakrishnan / Budhaditya Mukherjee / Ved Mehta / Matteo Lunghi / Jonas Grossmann / Bohumil Maco / Rémy Visentin / Adrian B Hehl / Volodymyr M Korkhov / Dominique Soldati-Favre / Abstract: Active host cell invasion by the obligate intracellular apicomplexan parasites relies on the formation of a moving junction, which connects parasite and host cell plasma membranes during entry. ...Active host cell invasion by the obligate intracellular apicomplexan parasites relies on the formation of a moving junction, which connects parasite and host cell plasma membranes during entry. Invading Toxoplasma gondii tachyzoites secrete their rhoptry content and insert a complex of RON proteins on the cytoplasmic side of the host cell membrane providing an anchor to which the parasite tethers. Here we show that a rhoptry-resident kinase RON13 is a key virulence factor that plays a crucial role in host cell entry. Cryo-EM, kinase assays, phosphoproteomics and cellular analyses reveal that RON13 is a secretory pathway kinase of atypical structure that phosphorylates rhoptry proteins including the components of the RON complex. Ultimately, RON13 kinase activity controls host cell invasion by anchoring the moving junction at the parasite-host cell interface. | |||||||||
History |
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-Structure visualization
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Structure viewer | Molecule: MolmilJmol/JSmol |
-Downloads & links
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PDBx/mmCIF format | 7nur.cif.gz | 191 KB | Display | PDBx/mmCIF format |
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PDB format | pdb7nur.ent.gz | 153.3 KB | Display | PDB format |
PDBx/mmJSON format | 7nur.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7nur_validation.pdf.gz | 453 KB | Display | wwPDB validaton report |
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Full document | 7nur_full_validation.pdf.gz | 460.8 KB | Display | |
Data in XML | 7nur_validation.xml.gz | 35.2 KB | Display | |
Data in CIF | 7nur_validation.cif.gz | 50.8 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/nu/7nur ftp://data.pdbj.org/pub/pdb/validation_reports/nu/7nur | HTTPS FTP |
-Related structure data
Related structure data | 12600MC M: map data used to model this data C: citing same article (ref.) |
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Similar structure data |
-Links
-Assembly
Deposited unit |
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1 |
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-Components
#1: Protein | Mass: 126419.680 Da / Num. of mol.: 1 / Mutation: D595A Source method: isolated from a genetically manipulated source Source: (gene. exp.) Toxoplasma gondii (strain ATCC 50853 / GT1) (eukaryote) Strain: ATCC 50853 / GT1 / Gene: TGGT1_321650 / Cell line (production host): Sf9 / Production host: Spodoptera frugiperda (fall armyworm) / References: UniProt: S7VRX1 |
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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: Secretory pathway kinase Ron13 / Type: ORGANELLE OR CELLULAR COMPONENT / Entity ID: all / Source: RECOMBINANT |
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Molecular weight | Experimental value: NO |
Source (natural) | Organism: Toxoplasma gondii (eukaryote) |
Source (recombinant) | Organism: Spodoptera frugiperda (fall armyworm) / Cell: Sf9 / Plasmid: pFastBac |
Buffer solution | pH: 7.5 Details: 5 mM Tris pH 7.5 / 200 mM NaCl / 0.5 mM EDTA / 3 mM DTT |
Specimen | Conc.: 15 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy imaging
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
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Microscopy | Model: FEI TITAN KRIOS |
Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
Electron lens | Mode: BRIGHT FIELD |
Image recording | Electron dose: 47 e/Å2 / Film or detector model: GATAN K2 QUANTUM (4k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
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Symmetry | Point symmetry: C1 (asymmetric) |
3D reconstruction | Resolution: 3.125 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 320723 / Symmetry type: POINT |