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Yorodumi- PDB-6ef3: Yeast 26S proteasome bound to ubiquitinated substrate (4D motor state) -
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Open data
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Basic information
| Entry | Database: PDB / ID: 6ef3 | |||||||||
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| Title | Yeast 26S proteasome bound to ubiquitinated substrate (4D motor state) | |||||||||
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Keywords | MOTOR PROTEIN / 26S Proteasome / ATPase / AAA+ / Protease / Ubiquitin | |||||||||
| Function / homology | Function and homology informationmitotic cell cycle phase transition / proteasome regulatory particle assembly / proteasome storage granule assembly / proteasome regulatory particle, base subcomplex / peroxisome fission / protein-containing complex localization / mitochondrial fission / proteasome-activating activity / proteasome regulatory particle, lid subcomplex / ER-Phagosome pathway ...mitotic cell cycle phase transition / proteasome regulatory particle assembly / proteasome storage granule assembly / proteasome regulatory particle, base subcomplex / peroxisome fission / protein-containing complex localization / mitochondrial fission / proteasome-activating activity / proteasome regulatory particle, lid subcomplex / ER-Phagosome pathway / Antigen processing: Ub, ATP-independent proteasomal degradation / 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 / Proteasome assembly / nonfunctional rRNA decay / Ub-specific processing proteases / metal-dependent deubiquitinase activity / peptide catabolic process / proteasome binding / proteasomal ubiquitin-independent protein catabolic process / positive regulation of RNA polymerase II transcription preinitiation complex assembly / ribosomal large subunit export from nucleus / proteasome storage granule / protein deubiquitination / proteasome endopeptidase complex / proteasome core complex, beta-subunit complex / endopeptidase activator activity / threonine-type endopeptidase activity / proteasome core complex, alpha-subunit complex / proteasome assembly / proteasome complex / ERAD pathway / Neutrophil degranulation / positive regulation of transcription elongation by RNA polymerase II / nucleotide-excision repair / modification-dependent protein catabolic process / metallopeptidase activity / protein tag activity / positive regulation of protein catabolic process / cytosolic ribosome / positive regulation of proteasomal ubiquitin-dependent protein catabolic process / peroxisome / ribosome biogenesis / ribosomal large subunit assembly / endopeptidase activity / ubiquitin-dependent protein catabolic process / cytosolic large ribosomal subunit / proteasome-mediated ubiquitin-dependent protein catabolic process / cytoplasmic translation / cell division / ubiquitinyl hydrolase 1 / cysteine-type deubiquitinase activity / protein ubiquitination / chromatin remodeling / structural constituent of ribosome / protein domain specific binding / ubiquitin protein ligase binding / mRNA binding / endoplasmic reticulum membrane / ATP hydrolysis activity / mitochondrion / ATP binding / identical protein binding / nucleus / cytosol / cytoplasm Similarity search - Function | |||||||||
| Biological species | ![]() Homo sapiens (human) | |||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.17 Å | |||||||||
Authors | de la Pena, A.H. / Goodall, E.A. / Gates, S.N. / Lander, G.C. / Martin, A. | |||||||||
| Funding support | United States, 2items
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Citation | Journal: Science / Year: 2018Title: 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. | |||||||||
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Structure visualization
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| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 6ef3.cif.gz | 1010.9 KB | Display | PDBx/mmCIF format |
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| PDB format | pdb6ef3.ent.gz | 804.3 KB | Display | PDB format |
| PDBx/mmJSON format | 6ef3.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/ef/6ef3 ftp://data.pdbj.org/pub/pdb/validation_reports/ef/6ef3 | HTTPS FTP |
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-Related structure data
| Related structure data | ![]() 9045MC ![]() 9042C ![]() 9043C ![]() 9044C ![]() 6ef0C ![]() 6ef1C ![]() 6ef2C M: map data used to model this data C: citing same article ( |
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| Similar structure data |
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Links
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Assembly
| Deposited unit | ![]()
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Components
-Proteasome subunit beta type- ... , 8 types, 8 molecules 1234567n
| #1: Protein | Mass: 23573.604 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P38624, proteasome endopeptidase complex |
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| #2: Protein | Mass: 28299.889 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P25043, proteasome endopeptidase complex |
| #3: Protein | Mass: 22627.842 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P25451, proteasome endopeptidase complex |
| #4: Protein | Mass: 22545.676 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P22141, proteasome endopeptidase complex |
| #5: Protein | Mass: 31670.539 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P30656, proteasome endopeptidase complex |
| #6: Protein | Mass: 26905.076 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P23724, proteasome endopeptidase complex |
| #7: Protein | Mass: 29471.289 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P30657, proteasome endopeptidase complex |
| #21: Protein/peptide | Mass: 1789.039 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
-Proteasome subunit alpha type- ... , 6 types, 6 molecules ABCDEF
| #8: Protein | Mass: 28033.830 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P21243, proteasome endopeptidase complex |
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| #9: Protein | Mass: 27191.828 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P23639, proteasome endopeptidase complex |
| #10: Protein | Mass: 28748.230 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P23638, proteasome endopeptidase complex |
| #11: Protein | Mass: 28478.111 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P40303, proteasome endopeptidase complex |
| #12: Protein | Mass: 28649.086 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P32379, proteasome endopeptidase complex |
| #13: Protein | Mass: 25634.000 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P40302, proteasome endopeptidase complex |
-Protein , 4 types, 4 molecules GLru
| #14: Protein | Mass: 31575.068 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() References: UniProt: P21242, proteasome endopeptidase complex |
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| #19: Protein | Mass: 49480.137 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
| #22: Protein | Mass: 34442.281 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
| #24: Protein | Mass: 14583.077 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
-26S proteasome regulatory subunit ... , 5 types, 5 molecules HIJKM
| #15: Protein | Mass: 52054.891 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
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| #16: Protein | Mass: 48898.160 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
| #17: Protein | Mass: 45342.742 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
| #18: Protein | Mass: 47953.676 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
| #20: Protein | Mass: 48315.727 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() |
-Protein/peptide , 1 types, 1 molecules s
| #23: Protein/peptide | Mass: 4068.414 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Production host: ![]() |
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-Non-polymers , 2 types, 6 molecules 


| #25: Chemical | ChemComp-ATP / #26: Chemical | |
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-Details
| Has protein modification | Y |
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-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
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| Buffer solution | pH: 7.6 | ||||||||||||||||||||||||||||||||||||
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| Specimen | Conc.: 25 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES 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 ...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 | ||||||||||||||||||||||||||||||||||||
| Specimen support | Details: unspecified | ||||||||||||||||||||||||||||||||||||
| Vitrification | Instrument: HOMEMADE PLUNGER / Cryogen name: ETHANE / Humidity: 90 % / Chamber temperature: 277 K Details: specimens were manually blotted with Whatman #1 filter paper |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TITAN KRIOS / Details: images were acquired in nanoprobe mode |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 29000 X / Nominal defocus max: -2500 nm / Nominal defocus min: -1000 nm / Calibrated defocus min: -1500 nm / Calibrated defocus max: -3000 nm / Cs: 2.7 mm / C2 aperture diameter: 70 µm / Alignment procedure: COMA FREE |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Average exposure time: 6.25 sec. / Electron dose: 50 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Num. of grids imaged: 1 / Num. of real images: 11656 |
| Image scans | Sampling size: 5 µm / Width: 3710 / Height: 3838 / Movie frames/image: 25 / Used frames/image: 1-25 |
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Processing
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| Image processing | Details: Camera was operated in counting mode | ||||||||||||||||||||||||||||||||||||||||||||||||
| CTF correction | Details: CTF correction was performed by Relion during reconstruction Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 579361 Details: Particles were selected using the Relion template-based particle picker | ||||||||||||||||||||||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 4.17 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 48335 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | Protocol: RIGID BODY FIT / Space: REAL | ||||||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | PDB-ID: 5MPC Accession code: 5MPC / Source name: PDB / Type: experimental model |
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About Yorodumi




Homo sapiens (human)
United States, 2items
Citation
UCSF Chimera












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