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Yorodumi- EMDB-12600: Structure of the Toxoplasma gondii kinase Ron13, kinase-dead mutant -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-12600 | |||||||||
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Title | Structure of the Toxoplasma gondii kinase Ron13, kinase-dead mutant | |||||||||
Map data | Ron13 cryo-EM map | |||||||||
Sample |
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Keywords | secretory pathway / kinase / TRANSFERASE | |||||||||
Function / homology | Function and homology information protein kinase activity / signal transduction / ATP binding / membrane / cytoplasm Similarity search - Function | |||||||||
Biological species | Toxoplasma gondii (eukaryote) / Toxoplasma gondii (strain ATCC 50853 / GT1) (eukaryote) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.125 Å | |||||||||
Authors | Korkhov VM / Mehta V | |||||||||
Funding support | Switzerland, 2 items
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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
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_12600.map.gz | 7.3 MB | EMDB map data format | |
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Header (meta data) | emd-12600-v30.xml emd-12600.xml | 11 KB 11 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_12600_fsc.xml | 10.7 KB | Display | FSC data file |
Images | emd_12600.png | 79.7 KB | ||
Filedesc metadata | emd-12600.cif.gz | 5.9 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-12600 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-12600 | HTTPS FTP |
-Validation report
Summary document | emd_12600_validation.pdf.gz | 378.6 KB | Display | EMDB validaton report |
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Full document | emd_12600_full_validation.pdf.gz | 378.1 KB | Display | |
Data in XML | emd_12600_validation.xml.gz | 11.9 KB | Display | |
Data in CIF | emd_12600_validation.cif.gz | 15.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12600 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-12600 | HTTPS FTP |
-Related structure data
Related structure data | 7nurMC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_12600.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Ron13 cryo-EM map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 0.8544 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 : Secretory pathway kinase Ron13
Entire | Name: Secretory pathway kinase Ron13 |
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Components |
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-Supramolecule #1: Secretory pathway kinase Ron13
Supramolecule | Name: Secretory pathway kinase Ron13 / type: organelle_or_cellular_component / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Toxoplasma gondii (eukaryote) |
-Macromolecule #1: Protein kinase domain-containing protein
Macromolecule | Name: Protein kinase domain-containing protein / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Toxoplasma gondii (strain ATCC 50853 / GT1) (eukaryote) Strain: ATCC 50853 / GT1 |
Molecular weight | Theoretical: 126.41968 KDa |
Recombinant expression | Organism: Spodoptera frugiperda (fall armyworm) |
Sequence | String: GAGTTGGKKD DQALSFLGDT KSASELNPPR LVCPDKPPTL PPREEQVRKA YALPLCELPW DDLGPMLGSG TFGRVYPLRR PACTEVTKG FVGRKFAVKI FWLKRKGMMN LFDTISQGGT PSAEQTDPGT IAAIKSEIRS LPTSSSAFRD MVRIADPTVD V EKIKGMAD ...String: GAGTTGGKKD DQALSFLGDT KSASELNPPR LVCPDKPPTL PPREEQVRKA YALPLCELPW DDLGPMLGSG TFGRVYPLRR PACTEVTKG FVGRKFAVKI FWLKRKGMMN LFDTISQGGT PSAEQTDPGT IAAIKSEIRS LPTSSSAFRD MVRIADPTVD V EKIKGMAD SLTVETIMKE AKTLRTVINT NGFYTEVGET GTIFTQMEKF VQAHRPEIWS TLSKASQEAQ ASKYAEIGLA DN HWSLPLA RVLVKDKNDV KHWALLIELF DGDLQPKTDK TGYSLDGWNA KSGGNVVLRE IFSSREALIG LTSKLVKPFV VMQ NLYSLG HFAIKPPNLL YKYFPGEKGR ASRLSVAAGD FGMAGLLHGD MILRGTLAFM APEMERVSGG LVAKPSYDVY ALAL TLASF WTAATELRDH YPWVEKCIKP TLKKMKDAPE FTFLRFASKT GPKLYEADTI YALSTCFAVG GKVEKLYHTG MPLLI RLKL SQMADPEPLA RVSMRHARFV FKAYAMLDKL LRAPQSEANA ETREEQLKQL QSLHIVQFLL FYLRMEPLTA ARDNTQ SYR RLARALLDFA RLDPVYQAAT ETVQPLPYEF FTEQKDWQNV KVEVSGSEVD ETIRKLRTSL TRDRSLSEDS WADLVDI MF GVSLDGLREV VTRVVYSRKT FLLEEKIGNA VKEAVAATYK FDPNTQLIAE DAPDRLFEVV RTDLGLSYPD DSELGRFL V HRVSKSHTAW ATVDRLARQA LRLALRREER TRQVYEQLLS GEKPSSESEK AFFDSVFSAV SVVSEANYFG LFWDFPSAG LFGVPPEEMQ AYVRKTHLAF VGKMWPVETQ KKILEAAVRV TVRGLNASLP ASLVDVYATV FAALPTKAPV SPPFLYGLER EEYSSLLFD AKLPEFKEMV AFWATRHELN IAVQTAVGKI PDATNLSDED IEKQLEGMLP AHLRSPSPAR FGWPPEAVAD N IRLFIREA KDELALHGPD MVHNRIRVNG RSKPPRRAAF LFHEIFRKAI AFKKDISVLQ FNQFFTDILK QSFDPQCRRF IA EVKKRVK SAPAEYVRVA DTEAVAPLFE GEGKDILKLV AVDPAARASD PEPNNCFLWT QAFLDDKTIV VSTGSSGSSG HHH HHHHHH H UniProtKB: Protein kinase domain-containing protein |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 15 mg/mL |
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Buffer | pH: 7.5 Details: 5 mM Tris pH 7.5 / 200 mM NaCl / 0.5 mM EDTA / 3 mM DTT |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Average electron dose: 47.0 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 |