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- EMDB-19898: Plasmodium falciparum sporozoite actin determined in situ -

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Basic information

Entry
Database: EMDB / ID: EMD-19898
TitlePlasmodium falciparum sporozoite actin determined in situ
Map data
Sample
  • Organelle or cellular component: F-actin
KeywordsF-actin / actin / glideosome / STRUCTURAL PROTEIN
Biological speciesPlasmodium falciparum (malaria parasite P. falciparum)
Methodsubtomogram averaging / cryo EM / Resolution: 28.0 Å
AuthorsPrazak V / Ferreira JL
Funding support France, United Kingdom, 2 items
OrganizationGrant numberCountry
Human Frontier Science Program (HFSP)LT000024/2020-L France
Wellcome Trust227774/Z/23/Z United Kingdom
CitationJournal: EMBO Rep / Year: 2025
Title: Molecular architecture of glideosome and nuclear F-actin in Plasmodium falciparum.
Authors: Vojtěch Pražák / Daven Vasishtan / Kay Grünewald / Ross G Douglas / Josie L Ferreira /
Abstract: Actin-based motility is required for the transmission of malaria sporozoites. While this has been shown biochemically, filamentous actin has remained elusive and has not been directly visualised ...Actin-based motility is required for the transmission of malaria sporozoites. While this has been shown biochemically, filamentous actin has remained elusive and has not been directly visualised inside the parasite. Using focused ion beam milling and electron cryo-tomography, we studied dynamic actin filaments in unperturbed Plasmodium falciparum cells for the first time. This allowed us to dissect the assembly, path and fate of actin filaments during parasite gliding and determine a complete 3D model of F-actin within sporozoites. We observe micrometre long actin filaments, much longer than expected from in vitro studies. After their assembly at the parasite's apical end, actin filaments continue to grow as they are transported down the cell as part of the glideosome machinery, and are disassembled at the basal end in a rate-limiting step. Large pores in the IMC, constrained to the basal end, may facilitate actin exchange between the pellicular space and cytosol for recycling and maintenance of directional flow. The data also reveal striking actin bundles in the nucleus. Implications for motility and transmission are discussed.
History
DepositionMar 20, 2024-
Header (metadata) releaseFeb 5, 2025-
Map releaseFeb 5, 2025-
UpdateMay 7, 2025-
Current statusMay 7, 2025Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_19898.png

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Map

FileDownload / File: emd_19898.map.gz / Format: CCP4 / Size: 251 KB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
Projections & slices

Image control

Size
Brightness
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AxesZ (Sec.)Y (Row.)X (Col.)
6.67 Å/pix.
x 40 pix.
= 266.88 Å		6.67 Å/pix.
x 40 pix.
= 266.88 Å		6.67 Å/pix.
x 40 pix.
= 266.88 Å

Surface

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Images are generated by Spider.

Voxel sizeX=Y=Z: 6.672 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 11.0
Minimum - Maximum-18.603418000000001 - 38.303294999999999
Average (Standard dev.)-2.9768035 (±5.13677)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions404040
Spacing404040
CellA=B=C: 266.88 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: Unfiltered map

Fileemd_19898_additional_1.map
AnnotationUnfiltered map
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Half map: half map 2

Fileemd_19898_half_map_1.map
Annotationhalf map 2
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Half map: half map 1

Fileemd_19898_half_map_2.map
Annotationhalf map 1
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AxesZYX

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Sample components

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Entire : F-actin

EntireName: F-actin
Components
  • Organelle or cellular component: F-actin

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Supramolecule #1: F-actin

SupramoleculeName: F-actin / type: organelle_or_cellular_component / ID: 1 / Parent: 0
Source (natural)Organism: Plasmodium falciparum (malaria parasite P. falciparum)

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Experimental details

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Structure determination

Methodcryo EM
Processingsubtomogram averaging
Aggregation statefilament

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Sample preparation

BufferpH: 7.5
GridModel: UltrAuFoil R1.2/1.3 / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 60 sec.
VitrificationCryogen name: ETHANE-PROPANE / Instrument: HOMEMADE PLUNGER
DetailsSporozoite stage parasites dissected from infected mosquito salivary glands were vitried on EM grids and FIB-milled. Actin was manually picked from celluar tomograms.

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 3.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 6.0 µm / Nominal defocus min: 3.0 µm
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionApplied symmetry - Helical parameters - Δz: 28.4 Å
Applied symmetry - Helical parameters - Δ&Phi: -166 °
Applied symmetry - Helical parameters - Axial symmetry: C1 (asymmetric)
Resolution.type: BY AUTHOR / Resolution: 28.0 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: PEET / Number subtomograms used: 11487
ExtractionNumber tomograms: 29 / Number images used: 21
CTF correctionType: PHASE FLIPPING ONLY
Final angle assignmentType: OTHER
FSC plot (resolution estimation)

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