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Yorodumi- EMDB-36598: Cryo-EM structure of human 26S proteasomal RP subcomplex (Ea stat... -
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Open data
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Basic information
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| Title | Cryo-EM structure of human 26S proteasomal RP subcomplex (Ea state) without any bound substrate. | |||||||||||||||
Map data | Sharpened cryo-EM map of Ea state of 26S RP (with single CP alpha ring) without substrate bound. | |||||||||||||||
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Keywords | protein degradation / macromolecular complex / ubiquitin-proteasome system / CYTOSOLIC PROTEIN | |||||||||||||||
| Function / homology | Function and homology informationthyrotropin-releasing hormone receptor binding / nuclear proteasome complex / host-mediated perturbation of viral transcription / positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / proteasome accessory complex / integrator complex / purine ribonucleoside triphosphate binding ...thyrotropin-releasing hormone receptor binding / nuclear proteasome complex / host-mediated perturbation of viral transcription / positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / proteasome accessory complex / integrator complex / purine ribonucleoside triphosphate binding / meiosis I / proteasome regulatory particle / cytosolic proteasome complex / positive regulation of proteasomal protein catabolic process / proteasome-activating activity / proteasome regulatory particle, lid subcomplex / proteasome regulatory particle, base subcomplex / metal-dependent deubiquitinase activity / protein K63-linked deubiquitination / negative regulation of programmed cell death / Regulation of ornithine decarboxylase (ODC) / Proteasome assembly / Cross-presentation of soluble exogenous antigens (endosomes) / proteasome core complex / Homologous DNA Pairing and Strand Exchange / Defective homologous recombination repair (HRR) due to BRCA1 loss of function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA1 binding function / Defective HDR through Homologous Recombination Repair (HRR) due to PALB2 loss of BRCA2/RAD51/RAD51C binding function / Resolution of D-loop Structures through Synthesis-Dependent Strand Annealing (SDSA) / Somitogenesis / Resolution of D-loop Structures through Holliday Junction Intermediates / K63-linked deubiquitinase activity / Impaired BRCA2 binding to RAD51 / proteasome binding / transcription factor binding / regulation of protein catabolic process / myofibril / proteasome storage granule / Presynaptic phase of homologous DNA pairing and strand exchange / general transcription initiation factor binding / blastocyst development / positive regulation of RNA polymerase II transcription preinitiation complex assembly / polyubiquitin modification-dependent protein binding / protein deubiquitination / immune system process / NF-kappaB binding / endopeptidase activator activity / mRNA export from nucleus / proteasome assembly / proteasome core complex, alpha-subunit complex / SARS-CoV-1 targets host intracellular signalling and regulatory pathways / enzyme regulator activity / regulation of proteasomal protein catabolic process / ERAD pathway / inclusion body / proteasome complex / TBP-class protein binding / proteolysis involved in protein catabolic process / sarcomere / Regulation of activated PAK-2p34 by proteasome mediated degradation / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / N-glycan trimming in the ER and Calnexin/Calreticulin cycle / Asymmetric localization of PCP proteins / Ubiquitin-dependent degradation of Cyclin D / SCF-beta-TrCP mediated degradation of Emi1 / NIK-->noncanonical NF-kB signaling / TNFR2 non-canonical NF-kB pathway / AUF1 (hnRNP D0) binds and destabilizes mRNA / stem cell differentiation / Vpu mediated degradation of CD4 / Assembly of the pre-replicative complex / Ubiquitin-Mediated Degradation of Phosphorylated Cdc25A / Degradation of DVL / Dectin-1 mediated noncanonical NF-kB signaling / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / negative regulation of inflammatory response to antigenic stimulus / Degradation of AXIN / P-body / Hh mutants are degraded by ERAD / lipopolysaccharide binding / Activation of NF-kappaB in B cells / Degradation of GLI1 by the proteasome / G2/M Checkpoints / Hedgehog ligand biogenesis / GSK3B and BTRC:CUL1-mediated-degradation of NFE2L2 / Defective CFTR causes cystic fibrosis / Autodegradation of the E3 ubiquitin ligase COP1 / Negative regulation of NOTCH4 signaling / Regulation of RUNX3 expression and activity / Vif-mediated degradation of APOBEC3G / Hedgehog 'on' state / Degradation of GLI2 by the proteasome / GLI3 is processed to GLI3R by the proteasome / FBXL7 down-regulates AURKA during mitotic entry and in early mitosis / APC/C:Cdh1 mediated degradation of Cdc20 and other APC/C:Cdh1 targeted proteins in late mitosis/early G1 / MAPK6/MAPK4 signaling / double-strand break repair via homologous recombination / Degradation of beta-catenin by the destruction complex / Oxygen-dependent proline hydroxylation of Hypoxia-inducible Factor Alpha Similarity search - Function | |||||||||||||||
| Biological species | Homo sapiens (human) | |||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||||||||
Authors | Hsu STD / Draczkowski P / Wang YS | |||||||||||||||
| Funding support | Taiwan, 4 items
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Citation | Journal: Nat Commun / Year: 2025Title: Structural basis of K11/K48-branched ubiquitin chain recognition by the human 26S proteasome. Authors: Piotr Draczkowski / Szu-Ni Chen / Ting Chen / Yong-Sheng Wang / Hsin-An Shih / Jessica Y C Huang / Ming-Chieh Tsai / Shu-Yu Lin / Steven Lin / Rosa Viner / Yuan-Chih Chang / Kuen-Phon Wu / ...Authors: Piotr Draczkowski / Szu-Ni Chen / Ting Chen / Yong-Sheng Wang / Hsin-An Shih / Jessica Y C Huang / Ming-Chieh Tsai / Shu-Yu Lin / Steven Lin / Rosa Viner / Yuan-Chih Chang / Kuen-Phon Wu / Shang-Te Danny Hsu / ![]() Abstract: Beyond the canonical K48-linked homotypic polyubiquitination for proteasome-targeted proteolysis, K11/K48-branched ubiquitin (Ub) chains are involved in fast-tracking protein turnover during cell ...Beyond the canonical K48-linked homotypic polyubiquitination for proteasome-targeted proteolysis, K11/K48-branched ubiquitin (Ub) chains are involved in fast-tracking protein turnover during cell cycle progression and proteotoxic stress. Here, we report cryo-EM structures of human 26S proteasome in a complex with a K11/K48-branched Ub chain. The structures revealed a multivalent substrate recognition mechanism involving a hitherto unknown K11-linked Ub binding site at the groove formed by RPN2 and RPN10 in addition to the canonical K48-linkage binding site formed by RPN10 and RPT4/5 coiled-coil. Additionally, RPN2 recognizes an alternating K11-K48-linkage through a conserved motif similar to the K48-specific T1 binding site of RPN1. The insights gleaned from these structures explain the molecular mechanism underlying the recognition of the K11/K48-branched Ub as a priority signal in the ubiquitin-mediated proteasomal degradation. | |||||||||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_36598.map.gz | 230.2 MB | EMDB map data format | |
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| Header (meta data) | emd-36598-v30.xml emd-36598.xml | 66.7 KB 66.7 KB | Display Display | EMDB header |
| Images | emd_36598.png | 144 KB | ||
| Filedesc metadata | emd-36598.cif.gz | 15.1 KB | ||
| Others | emd_36598_additional_1.map.gz emd_36598_half_map_1.map.gz emd_36598_half_map_2.map.gz | 122.9 MB 226.3 MB 226.3 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-36598 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-36598 | HTTPS FTP |
-Validation report
| Summary document | emd_36598_validation.pdf.gz | 969.3 KB | Display | EMDB validaton report |
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| Full document | emd_36598_full_validation.pdf.gz | 968.8 KB | Display | |
| Data in XML | emd_36598_validation.xml.gz | 15.8 KB | Display | |
| Data in CIF | emd_36598_validation.cif.gz | 18.8 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-36598 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-36598 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 8jriMC ![]() 8jrtC ![]() 8jtiC ![]() 8k0gC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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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_36598.map.gz / Format: CCP4 / Size: 244.1 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Sharpened cryo-EM map of Ea state of 26S RP (with single CP alpha ring) without substrate bound. | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.4 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Additional map: Unsharpened cryo-EM map of Ea state of 26S...
| File | emd_36598_additional_1.map | ||||||||||||
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| Annotation | Unsharpened cryo-EM map of Ea state of 26S RP (with single CP alpha ring) without substrate bound. | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: Cryo-EM half map A of Ea state of...
| File | emd_36598_half_map_1.map | ||||||||||||
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| Annotation | Cryo-EM half map A of Ea state of 26S RP (with single CP alpha ring) without substrate bound. | ||||||||||||
| Projections & Slices |
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| Density Histograms |
-Half map: Cryo-EM half map B of Ea state of...
| File | emd_36598_half_map_2.map | ||||||||||||
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| Annotation | Cryo-EM half map B of Ea state of 26S RP (with single CP alpha ring) without substrate bound. | ||||||||||||
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| Density Histograms |
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Sample components
+Entire : Human 26S proteasome
+Supramolecule #1: Human 26S proteasome
+Supramolecule #2: 19S regulatory particle (RP) of human 26S proteasome.
+Supramolecule #3: AAA+-ATPase of 19S regulatory particle (RP) of human 26S proteasome.
+Supramolecule #4: Alpha subunits belonging to the 20S core particle (CP) of human 2...
+Macromolecule #1: 26S protease regulatory subunit 7
+Macromolecule #2: 26S protease regulatory subunit 4
+Macromolecule #3: 26S protease regulatory subunit 8
+Macromolecule #4: 26S protease regulatory subunit 6B
+Macromolecule #5: Proteasome subunit alpha type-6
+Macromolecule #6: Proteasome subunit alpha type-2
+Macromolecule #7: Proteasome subunit alpha type-4
+Macromolecule #8: Proteasome subunit alpha type-7
+Macromolecule #9: Proteasome subunit alpha type-5
+Macromolecule #10: Proteasome subunit alpha type-1
+Macromolecule #11: Proteasome subunit alpha type-3
+Macromolecule #12: 26S proteasome non-ATPase regulatory subunit 1
+Macromolecule #13: 26S proteasome non-ATPase regulatory subunit 14
+Macromolecule #14: 26S proteasome non-ATPase regulatory subunit 3
+Macromolecule #15: 26S proteasome non-ATPase regulatory subunit 12
+Macromolecule #16: 26S proteasome non-ATPase regulatory subunit 11
+Macromolecule #17: 26S proteasome non-ATPase regulatory subunit 6
+Macromolecule #18: 26S proteasome non-ATPase regulatory subunit 7
+Macromolecule #19: 26S proteasome non-ATPase regulatory subunit 13
+Macromolecule #20: 26S proteasome non-ATPase regulatory subunit 4
+Macromolecule #21: 26S proteasome non-ATPase regulatory subunit 8
+Macromolecule #22: 26S proteasome complex subunit DSS1
+Macromolecule #23: 26S proteasome non-ATPase regulatory subunit 2
+Macromolecule #24: 26S protease regulatory subunit 10B
+Macromolecule #25: 26S protease regulatory subunit 6A
+Macromolecule #26: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #27: MAGNESIUM ION
+Macromolecule #28: 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: 7.6 Component:
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| Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR / Details: 20 mA | |||||||||||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV / Details: incubation time= 3 s blotting time= 2.5 s. | |||||||||||||||||||||
| Details | The complex was additionally supplemented with an excess of preformed and SEC-purified human RPN13:UCHL5 complex. |
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Electron microscopy
| Microscope | FEI TITAN KRIOS |
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| Specialist optics | Energy filter - Slit width: 20 eV |
| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Number grids imaged: 3 / Number real images: 15242 / Average exposure time: 1.8 sec. / Average electron dose: 49.0 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: 1.8 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 70000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model |
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| Refinement | Space: REAL / Protocol: OTHER / Overall B value: 88 / Target criteria: 0.8 |
| Output model | ![]() PDB-8jri: |
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About Yorodumi



Keywords
Homo sapiens (human)
Authors
Taiwan, 4 items
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Z (Sec.)
Y (Row.)
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FIELD EMISSION GUN

