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- EMDB-3606: Malaria-infected red blood cell section showing a segmented schizont -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-3606 | |||||||||
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Title | Malaria-infected red blood cell section showing a segmented schizont | |||||||||
![]() | Resin section tomogram of a malaria infected red blood cell showing a fully segmented schizont. | |||||||||
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Biological species | ![]() ![]() | |||||||||
Method | electron tomography / negative staining | |||||||||
![]() | Hale VL / Saibil HR | |||||||||
![]() | ![]() Title: Parasitophorous vacuole poration precedes its rupture and rapid host erythrocyte cytoskeleton collapse in egress. Authors: Victoria L Hale / Jean M Watermeyer / Fiona Hackett / Gema Vizcay-Barrena / Christiaan van Ooij / James A Thomas / Matthew C Spink / Maria Harkiolaki / Elizabeth Duke / Roland A Fleck / ...Authors: Victoria L Hale / Jean M Watermeyer / Fiona Hackett / Gema Vizcay-Barrena / Christiaan van Ooij / James A Thomas / Matthew C Spink / Maria Harkiolaki / Elizabeth Duke / Roland A Fleck / Michael J Blackman / Helen R Saibil / ![]() Abstract: In the asexual blood stages of malarial infection, merozoites invade erythrocytes and replicate within a parasitophorous vacuole to form daughter cells that eventually exit (egress) by sequential ...In the asexual blood stages of malarial infection, merozoites invade erythrocytes and replicate within a parasitophorous vacuole to form daughter cells that eventually exit (egress) by sequential rupture of the vacuole and erythrocyte membranes. The current model is that PKG, a malarial cGMP-dependent protein kinase, triggers egress, activating malarial proteases and other effectors. Using selective inhibitors of either PKG or cysteine proteases to separately inhibit the sequential steps in membrane perforation, combined with video microscopy, electron tomography, electron energy loss spectroscopy, and soft X-ray tomography of mature intracellular parasites, we resolve intermediate steps in egress. We show that the parasitophorous vacuole membrane (PVM) is permeabilized 10-30 min before its PKG-triggered breakdown into multilayered vesicles. Just before PVM breakdown, the host red cell undergoes an abrupt, dramatic shape change due to the sudden breakdown of the erythrocyte cytoskeleton, before permeabilization and eventual rupture of the erythrocyte membrane to release the parasites. In contrast to the previous view of PKG-triggered initiation of egress and a gradual dismantling of the host erythrocyte cytoskeleton over the course of schizont development, our findings identify an initial step in egress and show that host cell cytoskeleton breakdown is restricted to a narrow time window within the final stages of egress. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 43 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 10.3 KB 10.3 KB | Display Display | ![]() |
Images | ![]() | 281.8 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 3586C ![]() 3587C ![]() 3610C C: citing same article ( |
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EM raw data | ![]() Data size: 280.6 MB Data #1: Soft X-ray tomograms of Plasmodium falciparum infected human erythrocytes stalled in egress [Soft X-ray tomograms]) |
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Resin section tomogram of a malaria infected red blood cell showing a fully segmented schizont. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 16.25796 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Plasmodium falciparum infected human erythrocyte treated with the...
Entire | Name: Plasmodium falciparum infected human erythrocyte treated with the egress inhibitor Compound 1 |
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Components |
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-Supramolecule #1: Plasmodium falciparum infected human erythrocyte treated with the...
Supramolecule | Name: Plasmodium falciparum infected human erythrocyte treated with the egress inhibitor Compound 1 type: cell / ID: 1 / Parent: 0 Details: High pressure frozen and freeze-substituted cell section |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | negative staining |
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![]() | electron tomography |
Aggregation state | cell |
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Sample preparation
Buffer | pH: 7 / Details: RPMI medium |
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Staining | Type: NEGATIVE / Material: Uranyl Acetate |
Sugar embedding | Material: HM20 / Details: Freeze substitution |
Grid | Material: COPPER / Support film - Material: CARBON / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE |
High pressure freezing | Instrument: OTHER Details: The value given for _emd_high_pressure_freezing.instrument is Baltech HPM010. This is not in a list of allowed values set(['LEICA EM PACT2', 'LEICA EM PACT', 'EMS-002 RAPID IMMERSION ...Details: The value given for _emd_high_pressure_freezing.instrument is Baltech HPM010. This is not in a list of allowed values set(['LEICA EM PACT2', 'LEICA EM PACT', 'EMS-002 RAPID IMMERSION FREEZER', 'OTHER', 'LEICA EM HPM100', 'BAL-TEC HPM 010']) so OTHER is written into the XML file. |
Sectioning | Ultramicrotomy - Instrument: Leica EM UC7 / Ultramicrotomy - Temperature: 293 K / Ultramicrotomy - Final thickness: 100 |
Fiducial marker | Manufacturer: Electron Microscopy Sciences / Diameter: 10 nm |
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Electron microscopy
Microscope | FEI TECNAI 12 |
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Image recording | Film or detector model: GATAN ULTRASCAN 1000 (2k x 2k) / Average electron dose: 100.0 e/Å2 |
Electron beam | Acceleration voltage: 120 kV / Electron source: TUNGSTEN HAIRPIN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Sample stage | Specimen holder model: OTHER |
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Image processing
Final reconstruction | Algorithm: BACK PROJECTION / Software - Name: ![]() |
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