+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-29755 | |||||||||
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Title | Cryptosporidium parvum sporozoite basal end | |||||||||
Map data | Cryptosporidium parvum sporozoite basal end | |||||||||
Sample |
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Keywords | Conoid / Parasite / basal / CELL INVASION | |||||||||
Biological species | Cryptosporidium parvum Iowa (eukaryote) | |||||||||
Method | electron tomography / cryo EM | |||||||||
Authors | Martinez M / Chang Y-W / Mageswaran SK | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Nat Commun / Year: 2023 Title: Origin and arrangement of actin filaments for gliding motility in apicomplexan parasites revealed by cryo-electron tomography. Authors: Matthew Martinez / Shrawan Kumar Mageswaran / Amandine Guérin / William David Chen / Cameron Parker Thompson / Sabine Chavin / Dominique Soldati-Favre / Boris Striepen / Yi-Wei Chang / Abstract: The phylum Apicomplexa comprises important eukaryotic parasites that invade host tissues and cells using a unique mechanism of gliding motility. Gliding is powered by actomyosin motors that ...The phylum Apicomplexa comprises important eukaryotic parasites that invade host tissues and cells using a unique mechanism of gliding motility. Gliding is powered by actomyosin motors that translocate host-attached surface adhesins along the parasite cell body. Actin filaments (F-actin) generated by Formin1 play a central role in this critical parasitic activity. However, their subcellular origin, path and ultrastructural arrangement are poorly understood. Here we used cryo-electron tomography to image motile Cryptosporidium parvum sporozoites and reveal the cellular architecture of F-actin at nanometer-scale resolution. We demonstrate that F-actin nucleates at the apically positioned preconoidal rings and is channeled into the pellicular space between the parasite plasma membrane and the inner membrane complex in a conoid extrusion-dependent manner. Within the pellicular space, filaments on the inner membrane complex surface appear to guide the apico-basal flux of F-actin. F-actin concordantly accumulates at the basal end of the parasite. Finally, analyzing a Formin1-depleted Toxoplasma gondii mutant pinpoints the upper preconoidal ring as the conserved nucleation hub for F-actin in Cryptosporidium and Toxoplasma. Together, we provide an ultrastructural model for the life cycle of F-actin for apicomplexan gliding motility. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_29755.map.gz | 482.7 MB | EMDB map data format | |
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Header (meta data) | emd-29755-v30.xml emd-29755.xml | 10.7 KB 10.7 KB | Display Display | EMDB header |
Images | emd_29755.png | 176.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-29755 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-29755 | HTTPS FTP |
-Validation report
Summary document | emd_29755_validation.pdf.gz | 541.6 KB | Display | EMDB validaton report |
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Full document | emd_29755_full_validation.pdf.gz | 541.1 KB | Display | |
Data in XML | emd_29755_validation.xml.gz | 4.9 KB | Display | |
Data in CIF | emd_29755_validation.cif.gz | 5.9 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29755 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-29755 | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article (ref.) |
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-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_29755.map.gz / Format: CCP4 / Size: 842.9 MB / Type: IMAGE STORED AS SIGNED BYTE | ||||||||||||||||||||
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Annotation | Cryptosporidium parvum sporozoite basal end | ||||||||||||||||||||
Voxel size | X=Y=Z: 10.6 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
-Entire : Cryptosporidium parvum sporozoite basal end
Entire | Name: Cryptosporidium parvum sporozoite basal end |
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Components |
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-Supramolecule #1: Cryptosporidium parvum sporozoite basal end
Supramolecule | Name: Cryptosporidium parvum sporozoite basal end / type: cell / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Cryptosporidium parvum Iowa (eukaryote) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | electron tomography |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7.4 |
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Grid | Model: Quantifoil R2/2 / Material: COPPER / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 310 K / Instrument: LEICA EM GP / Details: Front blot for 4 seconds before plunging. |
Details | Excysted sporozoites from isolated oocysts |
Sectioning | Other: NO SECTIONING |
Fiducial marker | Manufacturer: Ted Pella, Inc / Diameter: 10 nm |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Phase plate: VOLTA PHASE PLATE / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Software | Name: SerialEM (ver. 3.8) |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number real images: 61 / Average exposure time: 0.4 sec. / Average electron dose: 2.3 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 4.0 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 33000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Details | Raw frames were gain normalized in serialEM and motion corrected using the alignframes command in IMOD |
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Final reconstruction | Algorithm: BACK PROJECTION / Software - Name: IMOD (ver. 4.11.5) / Number images used: 61 |