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Open data
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Basic information
Entry | ![]() | |||||||||
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Title | Preholo-Proteasome from Pre1-1 Pre4-1 Double Mutant | |||||||||
![]() | Final Map for Pre1-1 Pre4-1 double mutant preholo-proteasome | |||||||||
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![]() | Preholo-proteasome / ![]() ![]() | |||||||||
Function / homology | ![]() regulation of proteasome core complex assembly / proteasome core complex assembly / nuclear outer membrane-endoplasmic reticulum membrane network / Cross-presentation of soluble exogenous antigens (endosomes) / TNFR2 non-canonical NF-kB pathway / proteasomal ubiquitin-independent protein catabolic process / proteasome storage granule / endopeptidase activator activity / ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() Similarity search - Function | |||||||||
Biological species | ![]() ![]() ![]() ![]() ![]() ![]() | |||||||||
Method | ![]() ![]() | |||||||||
![]() | Walsh Jr RM / Rawson S / Schnell H / Velez B / Hanna J | |||||||||
Funding support | ![]()
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![]() | ![]() Title: Structure of the preholoproteasome reveals late steps in proteasome core particle biogenesis. Authors: Richard M Walsh / Shaun Rawson / Helena M Schnell / Benjamin Velez / Tamayanthi Rajakumar / John Hanna / ![]() Abstract: Assembly of the proteasome's core particle (CP), a barrel-shaped chamber of four stacked rings, requires five chaperones and five subunit propeptides. Fusion of two half-CP precursors yields a ...Assembly of the proteasome's core particle (CP), a barrel-shaped chamber of four stacked rings, requires five chaperones and five subunit propeptides. Fusion of two half-CP precursors yields a complete structure but remains immature until active site maturation. Here, using Saccharomyces cerevisiae, we report a high-resolution cryogenic electron microscopy structure of preholoproteasome, a post-fusion assembly intermediate. Our data reveal how CP midline-spanning interactions induce local changes in structure, facilitating maturation. Unexpectedly, we find that cleavage may not be sufficient for propeptide release, as residual interactions with chaperones such as Ump1 hold them in place. We evaluated previous models proposing that dynamic conformational changes in chaperones drive CP fusion and autocatalytic activation by comparing preholoproteasome to pre-fusion intermediates. Instead, the data suggest a scaffolding role for the chaperones Ump1 and Pba1/Pba2. Our data clarify key aspects of CP assembly, suggest that undiscovered mechanisms exist to explain CP fusion/activation, and have relevance for diseases of defective CP biogenesis. | |||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 168.2 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 36.7 KB 36.7 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 11.9 KB | Display | ![]() |
Images | ![]() | 62.2 KB | ||
Filedesc metadata | ![]() | 9.4 KB | ||
Others | ![]() ![]() | 165 MB 165 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8t08MC ![]() 8t0mC M: atomic model generated by this map C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
File | ![]() | ||||||||||||||||||||
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Annotation | Final Map for Pre1-1 Pre4-1 double mutant preholo-proteasome | ||||||||||||||||||||
Voxel size | X=Y=Z: 1.06 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: Half Map A for Pre1-1 Pre4-1 double mutant preholo-proteasome
File | emd_40938_half_map_1.map | ||||||||||||
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Annotation | Half Map A for Pre1-1 Pre4-1 double mutant preholo-proteasome | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Half Map B for Pre1-1 Pre4-1 double mutant preholo-proteasome
File | emd_40938_half_map_2.map | ||||||||||||
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Annotation | Half Map B for Pre1-1 Pre4-1 double mutant preholo-proteasome | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
+Entire : Preholo-Proteasome from Pre1-1 Pre4-1 Double Mutant
+Supramolecule #1: Preholo-Proteasome from Pre1-1 Pre4-1 Double Mutant
+Macromolecule #1: Proteasome subunit alpha type-1
+Macromolecule #2: Proteasome subunit alpha type-2
+Macromolecule #3: Proteasome subunit alpha type-3
+Macromolecule #4: Proteasome subunit alpha type-4
+Macromolecule #5: Proteasome subunit alpha type-5
+Macromolecule #6: Proteasome subunit alpha type-6
+Macromolecule #7: Proteasome subunit alpha type-7
+Macromolecule #8: Proteasome maturation factor UMP1
+Macromolecule #9: Proteasome subunit beta type-2
+Macromolecule #10: Proteasome subunit beta type-3
+Macromolecule #11: Proteasome subunit beta type-4
+Macromolecule #12: Proteasome subunit beta type-5
+Macromolecule #13: Proteasome subunit beta type-6
+Macromolecule #14: Proteasome subunit beta type-1
+Macromolecule #15: Proteasome chaperone 1
+Macromolecule #16: Proteasome assembly chaperone 2
+Macromolecule #17: Proteasome subunit beta type-7
-Experimental details
-Structure determination
Method | ![]() |
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Aggregation state | particle |
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Sample preparation
Concentration | 2.4 mg/mL | ||||||||||||
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Buffer | pH: 7.5 Component:
Details: Fluorinated Fos-Choline was added to the sample immediately prior to deposition on a grid for plunge freezing. | ||||||||||||
Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 400 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR / Pretreatment - Pressure: 0.039 kPa / Details: PELCO easiGLOW | ||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 295.15 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: ![]() |
Electron optics | C2 aperture diameter: 50.0 µm / Calibrated magnification: 47169 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD![]() |
Specialist optics | Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 25 eV |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 2 / Number real images: 33116 / Average exposure time: 3.8 sec. / Average electron dose: 54.3 e/Å2 |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: AB INITIO MODEL |
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Output model | ![]() PDB-8t08: |