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- EMDB-28126: Toxoplasma apical rings -

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Basic information

Entry
Database: EMDB / ID: EMD-28126
TitleToxoplasma apical rings
Map data
Sample
  • Organelle or cellular component: The apical rings of Toxoplasma gondii revealed by subtomogram averaging
KeywordsCryo-electron tomography / Subtomogram averaging / Apical ring / Toxoplasma gondii / CELL INVASION
Biological speciesToxoplasma gondii RH (eukaryote)
Methodsubtomogram averaging / cryo EM / Resolution: 59.67 Å
AuthorsSun SY / Pintilie GD
Funding support United States, 2 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01GM079429 United States
National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)DP2HL137186 United States
CitationJournal: mBio / Year: 2024
Title: Cryogenic electron tomography reveals novel structures in the apical complex of .
Authors: Stella Y Sun / Li-Av Segev-Zarko / Grigore D Pintilie / Chi Yong Kim / Sophia R Staggers / Michael F Schmid / Elizabeth S Egan / Wah Chiu / John C Boothroyd /
Abstract: Intracellular infectious agents, like the malaria parasite, , face the daunting challenge of how to invade a host cell. This problem may be even harder when the host cell in question is the ...Intracellular infectious agents, like the malaria parasite, , face the daunting challenge of how to invade a host cell. This problem may be even harder when the host cell in question is the enucleated red blood cell, which lacks the host machinery co-opted by many pathogens for internalization. Evolution has provided and related single-celled parasites within the phylum Apicomplexa with a collection of organelles at their apical end that mediate invasion. This apical complex includes at least two sets of secretory organelles, micronemes and rhoptries, and several structural features like apical rings and a putative pore through which proteins may be introduced into the host cell during invasion. We perform cryogenic electron tomography (cryo-ET) equipped with Volta Phase Plate on isolated and vitrified merozoites to visualize the apical machinery. Through tomographic reconstruction of cellular compartments, we see new details of known structures like the rhoptry tip interacting directly with a rosette resembling the recently described rhoptry secretory apparatus (RSA) or with an apical vesicle docked beneath the RSA. Subtomogram averaging reveals that the apical rings have a fixed number of repeating units, each of which is similar in overall size and shape to the units in the apical rings of tachyzoites of . Comparison of these polar rings in and parasites also reveals them to have a structurally conserved assembly pattern. These results provide new insight into the essential and structurally conserved features of this remarkable machinery used by apicomplexan parasites to invade their respective host cells.
IMPORTANCE: Malaria is an infectious disease caused by parasites of the genus and is a leading cause of morbidity and mortality globally. Upon infection, parasites invade and replicate in red blood ...IMPORTANCE: Malaria is an infectious disease caused by parasites of the genus and is a leading cause of morbidity and mortality globally. Upon infection, parasites invade and replicate in red blood cells, where they are largely protected from the immune system. To enter host cells, the parasites employ a specialized apparatus at their anterior end. In this study, advanced imaging techniques like cryogenic electron tomography (cryo-ET) and Volta Phase Plate enable unprecedented visualization of whole merozoites, revealing previously unknown structural details of their invasion machinery. Key findings include new insights into the structural conservation of apical rings shared between and its apicomplexan cousin, . These discoveries shed light on the essential and conserved elements of the invasion machinery used by these pathogens. Moreover, the research provides a foundation for understanding the molecular mechanisms underlying parasite-host interactions, potentially informing strategies for combating diseases caused by apicomplexan parasites.
History
DepositionSep 13, 2022-
Header (metadata) releaseOct 11, 2023-
Map releaseOct 11, 2023-
UpdateMar 20, 2024-
Current statusMar 20, 2024Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_28126.png

Downloads & links

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Map

FileDownload / File: emd_28126.map.gz / Format: CCP4 / Size: 15.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Voxel sizeX=Y=Z: 7.086 Å
Density
Contour LevelBy AUTHOR: 0.2
Minimum - Maximum-0.81857526 - 1.8924623
Average (Standard dev.)0.005457647 (±0.05892758)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions160160160
Spacing160160160
CellA=B=C: 1133.76 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: This map is obtained by positioning the reconstructed...

Fileemd_28126_additional_1.map
AnnotationThis map is obtained by positioning the reconstructed full map at multiple positions around the rings in tomograms. This map is cropped to show a small segment of the entire rings.
Projections & Slices
AxesZYX

Projections

Slices (1/2)
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Histogram

Histogram (log scale)

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Half map: #1

Fileemd_28126_half_map_1.map
Projections & Slices
AxesZYX

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Density Histograms

Histogram

Histogram (log scale)

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Half map: #2

Fileemd_28126_half_map_2.map
Projections & Slices
AxesZYX

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Slices (1/2)
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Histogram

Histogram (log scale)

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Sample components

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Entire : The apical rings of Toxoplasma gondii revealed by subtomogram ave...

EntireName: The apical rings of Toxoplasma gondii revealed by subtomogram averaging
Components
  • Organelle or cellular component: The apical rings of Toxoplasma gondii revealed by subtomogram averaging

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Supramolecule #1: The apical rings of Toxoplasma gondii revealed by subtomogram ave...

SupramoleculeName: The apical rings of Toxoplasma gondii revealed by subtomogram averaging
type: organelle_or_cellular_component / ID: 1 / Parent: 0
Source (natural)Organism: Toxoplasma gondii RH (eukaryote)

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation state3D array

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Sample preparation

BufferpH: 7
GridModel: EMS Lacey Carbon / Pretreatment - Type: PLASMA CLEANING
VitrificationCryogen name: ETHANE / Instrument: LEICA EM GP

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Electron microscopy

MicroscopeFEI TALOS ARCTICA
Electron beamAcceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 1.0010000000000001 µm / Nominal defocus min: 1.0 µm
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 1.8 e/Å2
Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company

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Image processing

ExtractionNumber tomograms: 59 / Number images used: 335
Final angle assignmentType: NOT APPLICABLE
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 59.67 Å / Resolution method: OTHER / Number subtomograms used: 335

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