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Yorodumi- EMDB-9259: The structure of the Plasmodium falciparum 20S proteasome in comp... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9259 | |||||||||
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Title | The structure of the Plasmodium falciparum 20S proteasome in complex with one PA28 activator. | |||||||||
Map data | sharpened map used for refinement | |||||||||
Sample |
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Keywords | proteasome / protease / HYDROLASE / 11S subunit / hydrolyse activator / proteasome activator / complex | |||||||||
Function / homology | Function and homology information Cross-presentation of soluble exogenous antigens (endosomes) / Orc1 removal from chromatin / CDK-mediated phosphorylation and removal of Cdc6 / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / KEAP1-NFE2L2 pathway / UCH proteinases / Ub-specific processing proteases / Neddylation / Antigen processing: Ubiquitination & Proteasome degradation / MAPK6/MAPK4 signaling ...Cross-presentation of soluble exogenous antigens (endosomes) / Orc1 removal from chromatin / CDK-mediated phosphorylation and removal of Cdc6 / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / KEAP1-NFE2L2 pathway / UCH proteinases / Ub-specific processing proteases / Neddylation / Antigen processing: Ubiquitination & Proteasome degradation / MAPK6/MAPK4 signaling / proteasome activator complex / ABC-family proteins mediated transport / AUF1 (hnRNP D0) binds and destabilizes mRNA / Neutrophil degranulation / proteasome core complex / proteasomal ubiquitin-independent protein catabolic process / regulation of G1/S transition of mitotic cell cycle / endopeptidase activator activity / proteasome endopeptidase complex / proteasome core complex, beta-subunit complex / proteasome core complex, alpha-subunit complex / threonine-type endopeptidase activity / proteasomal protein catabolic process / regulation of proteasomal protein catabolic process / ubiquitin-dependent protein catabolic process / proteasome-mediated ubiquitin-dependent protein catabolic process / endopeptidase activity / hydrolase activity / nucleoplasm / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Plasmodium falciparum 3D7 (eukaryote) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | |||||||||
Authors | Hanssen E / Xie SC / Leis A / Metcalfe RD / Gillett DL / Tilley L / Griffin MDW | |||||||||
Funding support | Australia, 2 items
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Citation | Journal: Nat Microbiol / Year: 2019 Title: The structure of the PA28-20S proteasome complex from Plasmodium falciparum and implications for proteostasis. Authors: Stanley C Xie / Riley D Metcalfe / Eric Hanssen / Tuo Yang / David L Gillett / Andrew P Leis / Craig J Morton / Michael J Kuiper / Michael W Parker / Natalie J Spillman / Wilson Wong / ...Authors: Stanley C Xie / Riley D Metcalfe / Eric Hanssen / Tuo Yang / David L Gillett / Andrew P Leis / Craig J Morton / Michael J Kuiper / Michael W Parker / Natalie J Spillman / Wilson Wong / Christopher Tsu / Lawrence R Dick / Michael D W Griffin / Leann Tilley / Abstract: The activity of the proteasome 20S catalytic core is regulated by protein complexes that bind to one or both ends. The PA28 regulator stimulates 20S proteasome peptidase activity in vitro, but its ...The activity of the proteasome 20S catalytic core is regulated by protein complexes that bind to one or both ends. The PA28 regulator stimulates 20S proteasome peptidase activity in vitro, but its role in vivo remains unclear. Here, we show that genetic deletion of the PA28 regulator from Plasmodium falciparum (Pf) renders malaria parasites more sensitive to the antimalarial drug dihydroartemisinin, indicating that PA28 may play a role in protection against proteotoxic stress. The crystal structure of PfPA28 reveals a bell-shaped molecule with an inner pore that has a strong segregation of charges. Small-angle X-ray scattering shows that disordered loops, which are not resolved in the crystal structure, extend from the PfPA28 heptamer and surround the pore. Using single particle cryo-electron microscopy, we solved the structure of Pf20S in complex with one and two regulatory PfPA28 caps at resolutions of 3.9 and 3.8 Å, respectively. PfPA28 binds Pf20S asymmetrically, strongly engaging subunits on only one side of the core. PfPA28 undergoes rigid body motions relative to Pf20S. Molecular dynamics simulations support conformational flexibility and a leaky interface. We propose lateral transfer of short peptides through the dynamic interface as a mechanism facilitating the release of proteasome degradation products. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_9259.map.gz | 142.8 MB | EMDB map data format | |
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Header (meta data) | emd-9259-v30.xml emd-9259.xml | 42.2 KB 42.2 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_9259_fsc.xml | 12.2 KB | Display | FSC data file |
Images | emd_9259.png | 99.6 KB | ||
Filedesc metadata | emd-9259.cif.gz | 9.8 KB | ||
Others | emd_9259_additional.map.gz emd_9259_half_map_1.map.gz emd_9259_half_map_2.map.gz | 122.2 MB 122.4 MB 122.3 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-9259 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-9259 | HTTPS FTP |
-Validation report
Summary document | emd_9259_validation.pdf.gz | 1017.5 KB | Display | EMDB validaton report |
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Full document | emd_9259_full_validation.pdf.gz | 1017.1 KB | Display | |
Data in XML | emd_9259_validation.xml.gz | 19.7 KB | Display | |
Data in CIF | emd_9259_validation.cif.gz | 25.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9259 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-9259 | HTTPS FTP |
-Related structure data
Related structure data | 6muxMC 9257C 9258C 6dfkC 6muvC 6muwC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_9259.map.gz / Format: CCP4 / Size: 155.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | sharpened map used for refinement | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.31 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: original unsharpened map
File | emd_9259_additional.map | ||||||||||||
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Annotation | original unsharpened map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Plasmodium falciparum 20S proteasome
File | emd_9259_half_map_1.map | ||||||||||||
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Annotation | Plasmodium falciparum 20S proteasome | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Plasmodium falciparum 20S proteasome
File | emd_9259_half_map_2.map | ||||||||||||
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Annotation | Plasmodium falciparum 20S proteasome | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : 20S proteasome/PA28 complex.
+Supramolecule #1: 20S proteasome/PA28 complex.
+Supramolecule #2: Plasmodium falciparum 20S proteasome
+Supramolecule #3: 11S activator of the Plasmodium falciparum proteasome.
+Macromolecule #1: 20S proteasome alpha-1 subunit
+Macromolecule #2: 20S proteasome alpha-2 subunit
+Macromolecule #3: 20S proteasome alpha-3 subunit
+Macromolecule #4: 20S proteasome alpha-4 subunit
+Macromolecule #5: 20S proteasome alpha-5 subunit
+Macromolecule #6: 20S proteasome alpha-6 subunit
+Macromolecule #7: 20S proteasome alpha-7 subunit
+Macromolecule #8: 20S proteasome beta-1 subunit
+Macromolecule #9: 20S proteasome beta-2 subunit
+Macromolecule #10: 20S proteasome beta-3 subunit
+Macromolecule #11: 20S proteasome beta-4 subunit
+Macromolecule #12: 20S proteasome beta-5 subunit
+Macromolecule #13: 20S proteasome beta-6 subunit
+Macromolecule #14: 20S proteasome beta-7 subunit
+Macromolecule #15: Proteasome activator PA28
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.7 mg/mL | |||||||||||||||
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Buffer | pH: 7.4 Component:
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Grid | Details: unspecified | |||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV / Details: wait time 0sec blot time 2sec blot force -1. |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Number real images: 5200 / Average exposure time: 10.0 sec. / Average electron dose: 32.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 100.0 µm / Calibrated defocus max: 3.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus min: 1.0 µm / Nominal magnification: 100000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Initial model |
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Refinement | Space: REAL / Protocol: AB INITIO MODEL / Overall B value: 82.68 | ||||||
Output model | PDB-6mux: |