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Yorodumi- EMDB-9045: Yeast 26S proteasome bound to ubiquitinated substrate (4D motor state) -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-9045 | |||||||||
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Title | Yeast 26S proteasome bound to ubiquitinated substrate (4D motor state) | |||||||||
Map data | Yeast 26S proteasome bound to ubiquitinated substrate (4D motor state) | |||||||||
Sample |
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Function / homology | Function and homology information peroxisome fission / proteasome storage granule assembly / proteasome regulatory particle assembly / nonfunctional rRNA decay / mitotic cell cycle phase transition / cytosolic proteasome complex / proteasome regulatory particle, lid subcomplex / proteasome-activating activity / protein-containing complex localization / mitochondrial fission ...peroxisome fission / proteasome storage granule assembly / proteasome regulatory particle assembly / nonfunctional rRNA decay / mitotic cell cycle phase transition / cytosolic proteasome complex / proteasome regulatory particle, lid subcomplex / proteasome-activating activity / protein-containing complex localization / mitochondrial fission / proteasome regulatory particle, base subcomplex / cyclin-dependent protein serine/threonine kinase regulator activity / 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 / Ub-specific processing proteases / peptide catabolic process / proteasome binding / protein deubiquitination / proteasome storage granule / endopeptidase activator activity / ribosomal large subunit export from nucleus / proteasome assembly / positive regulation of RNA polymerase II transcription preinitiation complex assembly / proteasome endopeptidase complex / proteasome core complex, beta-subunit complex / proteasome core complex, alpha-subunit complex / threonine-type endopeptidase activity / : / Neutrophil degranulation / proteasome complex / proteasomal protein catabolic process / nucleotide-excision repair / positive regulation of transcription elongation by RNA polymerase II / modification-dependent protein catabolic process / protein tag activity / ribosomal large subunit assembly / positive regulation of protein catabolic process / metallopeptidase activity / ribosome biogenesis / cytoplasmic translation / cytosolic large ribosomal subunit / ubiquitin-dependent protein catabolic process / proteasome-mediated ubiquitin-dependent protein catabolic process / endopeptidase activity / ubiquitinyl hydrolase 1 / cysteine-type deubiquitinase activity / protein ubiquitination / structural constituent of ribosome / chromatin remodeling / cell division / protein domain specific binding / mRNA binding / ubiquitin protein ligase binding / endoplasmic reticulum membrane / ATP hydrolysis activity / mitochondrion / ATP binding / identical protein binding / metal ion binding / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||
Biological species | Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast) / Homo sapiens (human) / Saccharomyces cerevisiae S288c (yeast) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.17 Å | |||||||||
Authors | de la Pena AH / Goodall EA / Gates SN / Lander GC / Martin A | |||||||||
Funding support | United States, 2 items
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Citation | Journal: Science / Year: 2018 Title: Substrate-engaged 26 proteasome structures reveal mechanisms for ATP-hydrolysis-driven translocation. Authors: Andres H de la Peña / Ellen A Goodall / Stephanie N Gates / Gabriel C Lander / Andreas Martin / Abstract: The 26 proteasome is the primary eukaryotic degradation machine and thus is critically involved in numerous cellular processes. The heterohexameric adenosine triphosphatase (ATPase) motor of the ...The 26 proteasome is the primary eukaryotic degradation machine and thus is critically involved in numerous cellular processes. The heterohexameric adenosine triphosphatase (ATPase) motor of the proteasome unfolds and translocates targeted protein substrates into the open gate of a proteolytic core while a proteasomal deubiquitinase concomitantly removes substrate-attached ubiquitin chains. However, the mechanisms by which ATP hydrolysis drives the conformational changes responsible for these processes have remained elusive. Here we present the cryo-electron microscopy structures of four distinct conformational states of the actively ATP-hydrolyzing, substrate-engaged 26 proteasome. These structures reveal how mechanical substrate translocation accelerates deubiquitination and how ATP-binding, -hydrolysis, and phosphate-release events are coordinated within the AAA+ (ATPases associated with diverse cellular activities) motor to induce conformational changes and propel the substrate through the central pore. | |||||||||
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
-Related structure data
Related structure data | 6ef3MC 9042C 9043C 9044C 6ef0C 6ef1C 6ef2C 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_9045.map.gz / Format: CCP4 / Size: 149.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Yeast 26S proteasome bound to ubiquitinated substrate (4D motor state) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.03 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
+Mask #1
+Mask #2
+Mask #3
+Mask #4
+Mask #5
+Mask #6
+Additional map: Global (sharpened)
+Additional map: Alpha ring (sharpened)
+Additional map: Alpha ring (half 1)
+Additional map: Alpha ring (half 2)
+Additional map: Beta ring (sharpened)
+Additional map: Beta ring (half 1)
+Additional map: Beta ring (half 2)
+Additional map: Motor (sharpened)
+Additional map: Motor (half 1)
+Additional map: Motor (half 2)
+Additional map: Global (half 1)
+Additional map: Global (half 2)
+Additional map: Rpn11 (sharpened)
+Additional map: Rpn11 (half 1)
+Additional map: Rpn11 (half 2)
+Additional map: N-ring (sharpened)
+Additional map: N-ring (half 1)
+Additional map: N-ring (half 2)
-Sample components
+Entire : Substrate-engaged 26S proteasome in the 4D state
+Supramolecule #1: Substrate-engaged 26S proteasome in the 4D state
+Supramolecule #2: Proteasome
+Supramolecule #3: substrate
+Macromolecule #1: Proteasome subunit beta type-1
+Macromolecule #2: Proteasome subunit beta type-2
+Macromolecule #3: Proteasome subunit beta type-3
+Macromolecule #4: Proteasome subunit beta type-4
+Macromolecule #5: Proteasome subunit beta type-5
+Macromolecule #6: Proteasome subunit beta type-6
+Macromolecule #7: Proteasome subunit beta type-7
+Macromolecule #8: Proteasome subunit alpha type-1
+Macromolecule #9: Proteasome subunit alpha type-2
+Macromolecule #10: Proteasome subunit alpha type-3
+Macromolecule #11: Proteasome subunit alpha type-4
+Macromolecule #12: Proteasome subunit alpha type-5
+Macromolecule #13: Proteasome subunit alpha type-6
+Macromolecule #14: Probable proteasome subunit alpha type-7
+Macromolecule #15: 26S proteasome regulatory subunit 7 homolog
+Macromolecule #16: 26S proteasome regulatory subunit 4 homolog
+Macromolecule #17: 26S proteasome regulatory subunit 8 homolog
+Macromolecule #18: 26S proteasome regulatory subunit 6B homolog
+Macromolecule #19: 26S proteasome subunit RPT4
+Macromolecule #20: 26S proteasome regulatory subunit 6A
+Macromolecule #21: Proteasome subunit beta type-7
+Macromolecule #22: Ubiquitin carboxyl-terminal hydrolase RPN11
+Macromolecule #23: Model substrate polypeptide
+Macromolecule #24: Ubiquitin-60S ribosomal protein L40
+Macromolecule #25: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #26: ADENOSINE-5'-DIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 25 mg/mL | ||||||||||||||||||
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Buffer | pH: 7.6 Component:
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Grid | Support film - Material: CARBON / Support film - topology: HOLEY / Details: unspecified | ||||||||||||||||||
Vitrification | Cryogen name: ETHANE / Chamber humidity: 90 % / Chamber temperature: 277 K / Instrument: HOMEMADE PLUNGER Details: specimens were manually blotted with Whatman #1 filter paper. | ||||||||||||||||||
Details | 26S proteasomes were diluted to a concentration of 20 micromolar in a buffer with an ATP regeneration system, and 6 mM ortho-phenanthroline. This solution was mixed with an equal volume of 50 micromolar ubiquitinated model substrate |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Calibrated defocus max: -3.0 µm / Calibrated defocus min: -1.5 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: -2.5 µm / Nominal defocus min: -1.0 µm / Nominal magnification: 29000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Details | images were acquired in nanoprobe mode |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3710 pixel / Digitization - Dimensions - Height: 3838 pixel / Digitization - Sampling interval: 5.0 µm / Digitization - Frames/image: 1-25 / Number grids imaged: 1 / Number real images: 11656 / Average exposure time: 6.25 sec. / Average electron dose: 50.0 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |