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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-31730 | |||||||||
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| Title | 20S proteasome (after post-processing) | |||||||||
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 Sample | 
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| Biological species |  Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.22 Å | |||||||||
 Authors | Xu C / Cong Y | |||||||||
| Funding support |   Israel, 1 items 
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 Citation |  Journal: Nat Commun / Year: 2021Title: The 20S as a stand-alone proteasome in cells can degrade the ubiquitin tag. Authors: Indrajit Sahu / Sachitanand M Mali / Prasad Sulkshane / Cong Xu / Andrey Rozenberg / Roni Morag / Manisha Priyadarsini Sahoo / Sumeet K Singh / Zhanyu Ding / Yifan Wang / Sharleen Day / Yao ...Authors: Indrajit Sahu / Sachitanand M Mali / Prasad Sulkshane / Cong Xu / Andrey Rozenberg / Roni Morag / Manisha Priyadarsini Sahoo / Sumeet K Singh / Zhanyu Ding / Yifan Wang / Sharleen Day / Yao Cong / Oded Kleifeld / Ashraf Brik / Michael H Glickman /     ![]() Abstract: The proteasome, the primary protease for ubiquitin-dependent proteolysis in eukaryotes, is usually found as a mixture of 30S, 26S, and 20S complexes. These complexes have common catalytic sites, ...The proteasome, the primary protease for ubiquitin-dependent proteolysis in eukaryotes, is usually found as a mixture of 30S, 26S, and 20S complexes. These complexes have common catalytic sites, which makes it challenging to determine their distinctive roles in intracellular proteolysis. Here, we chemically synthesize a panel of homogenous ubiquitinated proteins, and use them to compare 20S and 26S proteasomes with respect to substrate selection and peptide-product generation. We show that 20S proteasomes can degrade the ubiquitin tag along with the conjugated substrate. Ubiquitin remnants on branched peptide products identified by LC-MS/MS, and flexibility in the 20S gate observed by cryo-EM, reflect the ability of the 20S proteasome to proteolyze an isopeptide-linked ubiquitin-conjugate. Peptidomics identifies proteasome-trapped ubiquitin-derived peptides and peptides of potential 20S substrates in Hi20S cells, hypoxic cells, and human failing-heart. Moreover, elevated levels of 20S proteasomes appear to contribute to cell survival under stress associated with damaged proteins.  | |||||||||
| History | 
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Structure visualization
| Movie | 
 
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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_31730.map.gz | 4.4 MB |  EMDB map data format | |
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| Header (meta data) |  emd-31730-v30.xml emd-31730.xml | 9.6 KB 9.6 KB  | Display Display  |  EMDB header | 
| Images |  emd_31730.png | 26.1 KB | ||
| Archive directory |  http://ftp.pdbj.org/pub/emdb/structures/EMD-31730 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-31730 | HTTPS FTP  | 
-Validation report
| Summary document |  emd_31730_validation.pdf.gz | 329.7 KB | Display |  EMDB validaton report | 
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| Full document |  emd_31730_full_validation.pdf.gz | 329.3 KB | Display | |
| Data in XML |  emd_31730_validation.xml.gz | 5.9 KB | Display | |
| Data in CIF |  emd_31730_validation.cif.gz | 6.8 KB | Display | |
| Arichive directory |  https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-31730 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-31730 | HTTPS FTP  | 
-Related structure data
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Links
| EMDB pages |  EMDB (EBI/PDBe) /  EMDataResource | 
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Map
| File |  Download / File: emd_31730.map.gz / Format: CCP4 / Size: 40.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Projections & slices | Image control
 
 Images are generated by Spider.  | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.318 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 : 20S proteasome incubated with monoUb CyclinB1-NT
| Entire | Name: 20S proteasome incubated with monoUb CyclinB1-NT | 
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| Components | 
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-Supramolecule #1: 20S proteasome incubated with monoUb CyclinB1-NT
| Supramolecule | Name: 20S proteasome incubated with monoUb CyclinB1-NT / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#14 | 
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| Source (natural) | Organism:  Homo sapiens (human) | 
-Experimental details
-Structure determination
| Method | cryo EM | 
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 Processing | single particle reconstruction | 
| Aggregation state | particle | 
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Sample preparation
| Buffer | pH: 7.4 | 
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| Vitrification | Cryogen name: ETHANE | 
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Electron microscopy
| Microscope | FEI TITAN KRIOS | 
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 38.0 e/Å2 | 
| Electron beam | Acceleration voltage: 300 kV / Electron source:  FIELD EMISSION GUN | 
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD | 
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company  | 
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Image processing
| Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.22 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 154436 | 
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| Initial angle assignment | Type: MAXIMUM LIKELIHOOD | 
| Final angle assignment | Type: MAXIMUM LIKELIHOOD | 
Movie
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About Yorodumi



Homo sapiens (human)
Authors
Israel, 1 items 
Citation
 
UCSF Chimera














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