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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-3537 | ||||||||||||
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| Title | 26S proteasome in presence of BeFx (s4) | ||||||||||||
Map data | 26S proteasome in presence of BeFx (s4) | ||||||||||||
Sample |
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Keywords | Macromolecular complex / 26S proteasome / Protease / Hydrolase | ||||||||||||
| Function / homology | Function and homology informationnegative regulation of ERAD pathway / SAGA complex localization to transcription regulatory region / negative regulation of proteasomal protein catabolic process / mitochondria-associated ubiquitin-dependent protein catabolic process / regulation of proteasomal ubiquitin-dependent protein catabolic process / proteasome regulatory particle assembly / proteasome storage granule assembly / peroxisome fission / transcription export complex 2 / maintenance of DNA trinucleotide repeats ...negative regulation of ERAD pathway / SAGA complex localization to transcription regulatory region / negative regulation of proteasomal protein catabolic process / mitochondria-associated ubiquitin-dependent protein catabolic process / regulation of proteasomal ubiquitin-dependent protein catabolic process / proteasome regulatory particle assembly / proteasome storage granule assembly / peroxisome fission / transcription export complex 2 / maintenance of DNA trinucleotide repeats / protein deneddylation / filamentous growth / protein-containing complex localization / COP9 signalosome / mitochondrial fission / proteasome regulatory particle / proteasome-activating activity / proteasome regulatory particle, lid subcomplex / proteasome regulatory particle, base subcomplex / ER-Phagosome pathway / Antigen processing: Ub, ATP-independent proteasomal degradation / regulation of protein catabolic process / proteasome core complex assembly / K48-linked polyubiquitin modification-dependent protein binding / 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 / proteasome storage granule / polyubiquitin modification-dependent protein binding / proteasome endopeptidase complex / protein deubiquitination / proteasome core complex, beta-subunit complex / endopeptidase activator activity / threonine-type endopeptidase activity / proteasome core complex, alpha-subunit complex / proteasome assembly / mRNA export from nucleus / proteasome complex / enzyme regulator activity / ERAD pathway / Neutrophil degranulation / protein folding chaperone / ubiquitin binding / positive regulation of transcription elongation by RNA polymerase II / nucleotide-excision repair / double-strand break repair via homologous recombination / metallopeptidase activity / positive regulation of protein catabolic process / positive regulation of proteasomal ubiquitin-dependent protein catabolic process / peroxisome / endopeptidase activity / ubiquitin-dependent protein catabolic process / molecular adaptor activity / proteasome-mediated ubiquitin-dependent protein catabolic process / regulation of cell cycle / protein-macromolecule adaptor activity / ubiquitinyl hydrolase 1 / cysteine-type deubiquitinase activity / chromatin remodeling / protein domain specific binding / ubiquitin protein ligase binding / mRNA binding / endoplasmic reticulum membrane / structural molecule activity / endoplasmic reticulum / positive regulation of transcription by RNA polymerase II / ATP hydrolysis activity / mitochondrion / DNA-templated transcription / ATP binding / metal ion binding / identical protein binding / nucleus / cytosol / cytoplasm Similarity search - Function | ||||||||||||
| Biological species | ![]() ![]() | ||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 7.7 Å | ||||||||||||
Authors | Wehmer M / Rudack T | ||||||||||||
| Funding support | Germany, United States, 3 items
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Citation | Journal: Proc Natl Acad Sci U S A / Year: 2017Title: Structural insights into the functional cycle of the ATPase module of the 26S proteasome. Authors: Marc Wehmer / Till Rudack / Florian Beck / Antje Aufderheide / Günter Pfeifer / Jürgen M Plitzko / Friedrich Förster / Klaus Schulten / Wolfgang Baumeister / Eri Sakata / ![]() Abstract: In eukaryotic cells, the ubiquitin-proteasome system (UPS) is responsible for the regulated degradation of intracellular proteins. The 26S holocomplex comprises the core particle (CP), where ...In eukaryotic cells, the ubiquitin-proteasome system (UPS) is responsible for the regulated degradation of intracellular proteins. The 26S holocomplex comprises the core particle (CP), where proteolysis takes place, and one or two regulatory particles (RPs). The base of the RP is formed by a heterohexameric AAA ATPase module, which unfolds and translocates substrates into the CP. Applying single-particle cryo-electron microscopy (cryo-EM) and image classification to samples in the presence of different nucleotides and nucleotide analogs, we were able to observe four distinct conformational states (s1 to s4). The resolution of the four conformers allowed for the construction of atomic models of the AAA ATPase module as it progresses through the functional cycle. In a hitherto unobserved state (s4), the gate controlling access to the CP is open. The structures described in this study allow us to put forward a model for the 26S functional cycle driven by ATP hydrolysis. | ||||||||||||
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Structure visualization
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_3537.map.gz | 23.7 MB | EMDB map data format | |
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| Header (meta data) | emd-3537-v30.xml emd-3537.xml | 54.5 KB 54.5 KB | Display Display | EMDB header |
| Images | emd_3537.png | 54.6 KB | ||
| Filedesc metadata | emd-3537.cif.gz | 14.4 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-3537 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-3537 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 5mpcMC ![]() 3534C ![]() 3535C ![]() 3536C ![]() 5mp9C ![]() 5mpaC ![]() 5mpbC ![]() 5mpdC ![]() 5mpeC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_3537.map.gz / Format: CCP4 / Size: 274.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | 26S proteasome in presence of BeFx (s4) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.35 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 : 26S proteasome of Saccharomyces cerevisiae in presence of BeFx (s4)
+Supramolecule #1: 26S proteasome of Saccharomyces cerevisiae in presence of BeFx (s4)
+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: Probable proteasome subunit alpha type-7
+Macromolecule #8: Proteasome subunit beta type-1
+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-7
+Macromolecule #15: 26S protease regulatory subunit 7 homolog
+Macromolecule #16: 26S protease regulatory subunit 4 homolog
+Macromolecule #17: 26S protease regulatory subunit 6B homolog
+Macromolecule #18: 26S protease subunit RPT4
+Macromolecule #19: 26S protease regulatory subunit 6A
+Macromolecule #20: 26S protease regulatory subunit 8 homolog
+Macromolecule #21: 26S proteasome regulatory subunit RPN10
+Macromolecule #22: Ubiquitin carboxyl-terminal hydrolase RPN11
+Macromolecule #23: 26S proteasome regulatory subunit RPN12
+Macromolecule #24: 26S proteasome regulatory subunit RPN13
+Macromolecule #25: 26S proteasome complex subunit SEM1
+Macromolecule #26: 26S proteasome regulatory subunit RPN1
+Macromolecule #27: 26S proteasome regulatory subunit RPN2
+Macromolecule #28: 26S proteasome regulatory subunit RPN3
+Macromolecule #29: 26S proteasome regulatory subunit RPN5
+Macromolecule #30: 26S proteasome regulatory subunit RPN6
+Macromolecule #31: 26S proteasome regulatory subunit RPN7
+Macromolecule #32: 26S proteasome regulatory subunit RPN8
+Macromolecule #33: 26S proteasome regulatory subunit RPN9
+Macromolecule #34: Ubiquitin carboxyl-terminal hydrolase 6
+Macromolecule #35: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #36: MAGNESIUM ION
+Macromolecule #37: ADENOSINE-5'-DIPHOSPHATE
-Experimental details
-Structure determination
| Method | cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Concentration | 1 mg/mL |
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| Buffer | pH: 7.4 |
| Grid | Model: Quantifoil R2/1 / Material: COPPER / Mesh: 400 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 15 sec. |
| Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 95 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Image recording | Film or detector model: FEI FALCON II (4k x 4k) / Average electron dose: 45.0 e/Å2 |
| Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.5 µm |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
| Startup model | Type of model: PDB ENTRY PDB model - PDB ID: |
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| Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 7.7 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 1.4) / Number images used: 27600 |
| Initial angle assignment | Type: PROJECTION MATCHING / Software - Name: RELION (ver. 1.4) |
| Final angle assignment | Type: PROJECTION MATCHING / Software - Name: RELION (ver. 1.4) |
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Keywords
Authors
Germany,
United States, 3 items
Citation
UCSF Chimera





































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