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- PDB-7nur: Structure of the Toxoplasma gondii kinase Ron13, kinase-dead mutant -

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Basic information

Entry
Database: PDB / ID: 7nur
TitleStructure of the Toxoplasma gondii kinase Ron13, kinase-dead mutant
ComponentsProtein kinase domain-containing protein
KeywordsTRANSFERASE / secretory pathway / kinase
Function / homology
Function and homology information


protein kinase activity / ATP binding / membrane
Similarity search - Function
Serine/threonine-protein kinase, active site / Serine/Threonine protein kinases active-site signature. / Serine/Threonine protein kinases, catalytic domain / Protein kinase, ATP binding site / Protein kinases ATP-binding region signature. / Protein kinase domain profile. / Protein kinase domain / Protein kinase-like domain superfamily
Similarity search - Domain/homology
Protein kinase domain-containing protein
Similarity search - Component
Biological speciesToxoplasma gondii (eukaryote)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.125 Å
AuthorsKorkhov, V.M. / Mehta, V.
Funding support Switzerland, 2items
OrganizationGrant numberCountry
Swiss National Science Foundation310030B_166678 Switzerland
Swiss National Science Foundation310030_185325 Switzerland
CitationJournal: 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
DepositionMar 13, 2021Deposition site: PDBE / Processing site: PDBE
Revision 1.0May 26, 2021Provider: repository / Type: Initial release
Revision 1.1Jul 7, 2021Group: Database references / Category: citation / citation_author
Item: _citation.country / _citation.journal_abbrev ..._citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.page_first / _citation.page_last / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year / _citation_author.identifier_ORCID / _citation_author.name
Revision 1.2Oct 16, 2024Group: Data collection / Database references / Structure summary
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / em_admin / pdbx_entry_details / pdbx_modification_feature
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _em_admin.last_update

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Assembly

Deposited unit
A: Protein kinase domain-containing protein


Theoretical massNumber of molelcules
Total (without water)126,4201
Polymers126,4201
Non-polymers00
Water00
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration
TypeNameSymmetry operationNumber
identity operation1_5551
Buried area0 Å2
ΔGint0 kcal/mol
Surface area49460 Å2
MethodPISA

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Components

#1: Protein Protein kinase domain-containing 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
Has protein modificationY

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: Secretory pathway kinase Ron13 / Type: ORGANELLE OR CELLULAR COMPONENT / Entity ID: all / Source: RECOMBINANT
Molecular weightExperimental value: NO
Source (natural)Organism: Toxoplasma gondii (eukaryote)
Source (recombinant)Organism: Spodoptera frugiperda (fall armyworm) / Cell: Sf9 / Plasmid: pFastBac
Buffer solutionpH: 7.5
Details: 5 mM Tris pH 7.5 / 200 mM NaCl / 0.5 mM EDTA / 3 mM DTT
SpecimenConc.: 15 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

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Electron microscopy imaging

Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company
MicroscopyModel: FEI TITAN KRIOS
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELD
Image recordingElectron dose: 47 e/Å2 / Film or detector model: GATAN K2 QUANTUM (4k x 4k)

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Processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
SymmetryPoint symmetry: C1 (asymmetric)
3D reconstructionResolution: 3.125 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 320723 / Symmetry type: POINT

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