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- EMDB-26747: Plasmodium falciparum merozoite with a fused rhoptry pair -

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Basic information

Entry
Database: EMDB / ID: EMD-26747
TitlePlasmodium falciparum merozoite with a fused rhoptry pair
Map dataPlasmodium falciparum merozoite with a fused rhoptry pair
Sample
  • Cell: Plasmodium falciparum merozoite apical complex
KeywordsRhoptry / Parasite / CELL INVASION
Biological speciesPlasmodium falciparum 3D7 (eukaryote)
Methodelectron tomography / cryo EM
AuthorsMartinez M / Chang Y-W
Funding support United States, 1 items
OrganizationGrant numberCountry
David and Lucile Packard Foundation United States
CitationJournal: Nat Microbiol / Year: 2022
Title: Rhoptry secretion system structure and priming in Plasmodium falciparum revealed using in situ cryo-electron tomography.
Authors: Matthew Martinez / William David Chen / Marta Mendonça Cova / Petra Molnár / Shrawan Kumar Mageswaran / Amandine Guérin / Audrey R Odom John / Maryse Lebrun / Yi-Wei Chang /
Abstract: Apicomplexan parasites secrete contents of the rhoptries, club-shaped organelles in the apical region, into host cells to permit their invasion and establishment of infection. The rhoptry secretory ...Apicomplexan parasites secrete contents of the rhoptries, club-shaped organelles in the apical region, into host cells to permit their invasion and establishment of infection. The rhoptry secretory apparatus (RSA), which is critical for rhoptry secretion, was recently discovered in Toxoplasma and Cryptosporidium. It is unknown whether a similar molecular machinery exists in the malaria parasite Plasmodium. In this study, we use in situ cryo-electron tomography to investigate the rhoptry secretion system in P. falciparum merozoites. We identify the presence of an RSA at the cell apex and a morphologically distinct apical vesicle docking the tips of the two rhoptries to the RSA. We also discover two additional rhoptry organizations that lack the apical vesicle. Using subtomogram averaging, we reveal different conformations of the RSA structure corresponding to different rhoptry organizations. Our results highlight previously unknown steps in the process of rhoptry secretion and indicate a regulatory role for the conserved apical vesicle in host invasion by apicomplexan parasites.
History
DepositionApr 26, 2022-
Header (metadata) releaseMay 11, 2022-
Map releaseMay 11, 2022-
UpdateJan 17, 2024-
Current statusJan 17, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_26747.png

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Map

FileDownload / File: emd_26747.map.gz / Format: CCP4 / Size: 2.2 GB / Type: IMAGE STORED AS SIGNED INTEGER (2 BYTES)
AnnotationPlasmodium falciparum merozoite with a fused rhoptry pair
Voxel sizeX=Y=Z: 10.6 Å
Density
Minimum - Maximum-2002.0 - 1163.0
Average (Standard dev.)73.778809999999993 (±88.485245000000006)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin-209209-401
Dimensions14401023800
Spacing10231440800
CellA: 10843.801 Å / B: 15264.001 Å / C: 8480.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Plasmodium falciparum merozoite apical complex

EntireName: Plasmodium falciparum merozoite apical complex
Components
  • Cell: Plasmodium falciparum merozoite apical complex

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Supramolecule #1: Plasmodium falciparum merozoite apical complex

SupramoleculeName: Plasmodium falciparum merozoite apical complex / type: cell / ID: 1 / Parent: 0
Details: Two separate rhoptries lacking an associated apical vesicle
Source (natural)Organism: Plasmodium falciparum 3D7 (eukaryote)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7.4
Details: RPMI-1640 supplemented with 27 mM NaHCO3, 11 mM glucose, 5 mM HEPES, 0.01 mM thymidine, 1 mM sodium pyruvate, 0.37 mM hypoxanthine, 10 ug/mL gentamicin
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 297 K / Instrument: LEICA EM GP / Details: Front blot for 4-6 seconds before plunging.
DetailsMechanically isolated merozoites from synchronized schizonts
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: Ted Pella, Inc / Diameter: 10 nm

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 5.0 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 33000
Specialist opticsPhase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average exposure time: 0.5 sec. / Average electron dose: 140.0 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionAlgorithm: BACK PROJECTION / Software - Name: IMOD / Number images used: 61
DetailsRaw frames were gain normalized in serialEM and motion corrected using the alignframes command in IMOD

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