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- EMDB-74069: Rhoptry secretory apparatus of Toxoplasma gondii conformation 4 -

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Basic information

Entry
Database: EMDB / ID: EMD-74069
TitleRhoptry secretory apparatus of Toxoplasma gondii conformation 4
Map data
Sample
  • Cell: Nd-GTPase-KO Toxoplasma gondii tachyzoite
    • Complex: Rhoptry secretory apparatus
    • Organelle or cellular component: Apical vesicle
KeywordsRhoptry Secretion System / CELL INVASION
Biological speciesToxoplasma gondii (eukaryote)
Methodsubtomogram averaging / cryo EM / Resolution: 38.0 Å
AuthorsMorley L / Chang Y-W
Funding support United States, 4 items
OrganizationGrant numberCountry
David and Lucile Packard Foundation2019-69645 United States
Burroughs Wellcome Fund1022785 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R35 GM156396 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)T32 GM132039 United States
CitationJournal: Nat Microbiol / Year: 2026
Title: A conserved GTPase complex gates exocytosis for defence and parasitism across alveolates.
Authors: Marta Mendonça Cova / Lucas Morley / Marjorie Maynadier / Jason Delabre / Arnault Graindorge / Laurence Berry-Sterkers / Shrawan Kumar Mageswaran / Valérie Rofidal / Cameron J Powell / ...Authors: Marta Mendonça Cova / Lucas Morley / Marjorie Maynadier / Jason Delabre / Arnault Graindorge / Laurence Berry-Sterkers / Shrawan Kumar Mageswaran / Valérie Rofidal / Cameron J Powell / Martin J Boulanger / Yi-Wei Chang / Daniela Sparvoli / Maryse Lebrun /
Abstract: Regulated protein secretion is a fundamental process by which eukaryotic cells respond to and interact with their dynamic environments, yet its mechanism outside metazoans remains largely unknown. ...Regulated protein secretion is a fundamental process by which eukaryotic cells respond to and interact with their dynamic environments, yet its mechanism outside metazoans remains largely unknown. Here we uncover an atypical small GTPase complex (Nd-GTPase-Nd9-NdP1) conserved across the Alveolata-a major eukaryotic clade encompassing ciliates and apicomplexan parasites-that gates organelle exocytosis for diverse functions ranging from cellular defence to host invasion. Using Tetrahymena thermophila and Toxoplasma gondii as comparative models, we combine functional genetics, cryo-electron tomography and integrative structural modelling to show that this complex maintains the organelle secretory apparatus in a pre-secretion state and permits activation upon stimulation. Comparative analysis further reveals that apicomplexan parasites rely on an apical vesicle to tighten control over secretion. These findings define a unifying principle of gated alveolate exocytosis, illustrating how evolution remodels an ancient secretion system to meet specialized cellular demands, and underlie key mechanisms of parasite virulence.
History
DepositionNov 26, 2025-
Header (metadata) releaseOct 14, 2026-
Map releaseOct 14, 2026-
UpdateOct 14, 2026-
Current statusOct 14, 2026Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_74069.png

Downloads & links

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Map

FileDownload / File: emd_74069.map.gz / Format: CCP4 / Size: 3.8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
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AxesZ (Sec.)Y (Row.)X (Col.)
10 Å/pix.
x 100 pix.
= 1000. Å		10 Å/pix.
x 100 pix.
= 1000. Å		10 Å/pix.
x 100 pix.
= 1000. Å

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Images are generated by Spider.

Voxel sizeX=Y=Z: 10 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.73
Minimum - Maximum-16.654264000000001 - 20.712606000000001
Average (Standard dev.)0.00000003041303 (±0.8924057)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions100100100
Spacing100100100
CellA=B=C: 1000.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Half map: #1

Fileemd_74069_half_map_1.map
Projections & Slices
AxesZYX

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Density Histograms

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Half map: #2

Fileemd_74069_half_map_2.map
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Sample components

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Entire : Nd-GTPase-KO Toxoplasma gondii tachyzoite

EntireName: Nd-GTPase-KO Toxoplasma gondii tachyzoite
Components
  • Cell: Nd-GTPase-KO Toxoplasma gondii tachyzoite
    • Complex: Rhoptry secretory apparatus
    • Organelle or cellular component: Apical vesicle

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Supramolecule #1: Nd-GTPase-KO Toxoplasma gondii tachyzoite

SupramoleculeName: Nd-GTPase-KO Toxoplasma gondii tachyzoite / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Toxoplasma gondii (eukaryote) / Strain: RH Tati DeltaKu80

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Supramolecule #2: Rhoptry secretory apparatus

SupramoleculeName: Rhoptry secretory apparatus / type: complex / ID: 2 / Parent: 1
Details: 8-fold symmetric protein structure, anchored into the plasma membrane, which docks the associated apical vesicle to the plasma membrane
Source (natural)Organism: Toxoplasma gondii (eukaryote) / Strain: RH Tati DeltaKu80

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Supramolecule #3: Apical vesicle

SupramoleculeName: Apical vesicle / type: organelle_or_cellular_component / ID: 3 / Parent: 1
Details: A small vesicle at the rhoptry tips that is docked at the rhoptry secretory apparatus
Source (natural)Organism: Toxoplasma gondii (eukaryote) / Strain: RH Tati DeltaKu80

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statecell

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Sample preparation

BufferpH: 7.4
GridModel: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE
VitrificationCryogen name: ETHANE-PROPANE / Instrument: LEICA EM GP / Details: 4-6 seconds front blotting.

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Electron microscopy

MicroscopeTFS KRIOS
Specialist opticsPhase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 2.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 3.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 33000
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionNumber classes used: 2 / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 38.0 Å / Resolution method: OTHER / Software - Name: Dynamo / Details: Resolution estimated using ResMap / Number subtomograms used: 61
ExtractionNumber tomograms: 61 / Number images used: 61 / Method: Volumes picked manually / Software: (Name: IMOD, Dynamo, Warp)
CTF correctionSoftware - Name: Warp / Type: PHASE FLIPPING ONLY
Final 3D classificationNumber classes: 2 / Software - Name: Dynamo
Final angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: Dynamo

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