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Yorodumi- EMDB-47722: Nub1/Fat10-processing human 26S proteasome bound to Txnl1 with Rp... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-47722 | |||||||||
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Title | Nub1/Fat10-processing human 26S proteasome bound to Txnl1 with Rpt1 at top of spiral staircase | |||||||||
Map data | ||||||||||
Sample |
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Keywords | 26S Proteasome / Nub1 / Fat10 / MOTOR PROTEIN / HYDROLASE-PROTEIN BINDING complex | |||||||||
Function / homology | Function and homology information regulation of ubiquitin-dependent protein catabolic process / positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / disulfide oxidoreductase activity / modulation by host of viral transcription / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / proteasome accessory complex / integrator complex ...regulation of ubiquitin-dependent protein catabolic process / positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / disulfide oxidoreductase activity / modulation by host of viral transcription / 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 regulatory particle, lid subcomplex / proteasome-activating activity / proteasome regulatory particle, base subcomplex / metal-dependent deubiquitinase activity / negative regulation of programmed cell death / protein K63-linked deubiquitination / regulation of endopeptidase activity / 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) / RHOBTB1 GTPase cycle / Regulation of ornithine decarboxylase (ODC) / RND1 GTPase cycle / proteasome core complex / RND2 GTPase cycle / Resolution of D-loop Structures through Holliday Junction Intermediates / RND3 GTPase cycle / Cross-presentation of soluble exogenous antigens (endosomes) / K63-linked deubiquitinase activity / Somitogenesis / Impaired BRCA2 binding to RAD51 / RHOV GTPase cycle / myofibril / proteasome binding / immune system process / transcription factor binding / regulation of protein catabolic process / proteasome storage granule / Presynaptic phase of homologous DNA pairing and strand exchange / RHOU GTPase cycle / blastocyst development / protein-disulfide reductase activity / general transcription initiation factor binding / response to type II interferon / response to tumor necrosis factor / polyubiquitin modification-dependent protein binding / NF-kappaB binding / endopeptidase activator activity / proteasome assembly / protein deubiquitination / positive regulation of RNA polymerase II transcription preinitiation complex assembly / RHOBTB2 GTPase cycle / proteasome core complex, alpha-subunit complex / mRNA export from nucleus / SARS-CoV-1 targets host intracellular signalling and regulatory pathways / regulation of proteasomal protein catabolic process / enzyme regulator activity / ERAD pathway / inclusion body / sarcomere / proteasome complex / Regulation of activated PAK-2p34 by proteasome mediated degradation / proteolysis involved in protein catabolic process / N-glycan trimming in the ER and Calnexin/Calreticulin cycle / Autodegradation of Cdh1 by Cdh1:APC/C / APC/C:Cdc20 mediated degradation of Securin / Asymmetric localization of PCP proteins / SCF-beta-TrCP mediated degradation of Emi1 / Ubiquitin-dependent degradation of Cyclin D / NIK-->noncanonical NF-kB signaling / stem cell differentiation / TNFR2 non-canonical NF-kB pathway / AUF1 (hnRNP D0) binds and destabilizes mRNA / Vpu mediated degradation of CD4 / Assembly of the pre-replicative complex / Degradation of DVL / Ubiquitin Mediated Degradation of Phosphorylated Cdc25A / Cdc20:Phospho-APC/C mediated degradation of Cyclin A / Dectin-1 mediated noncanonical NF-kB signaling / negative regulation of inflammatory response to antigenic stimulus / lipopolysaccharide binding / Hh mutants are degraded by ERAD / Degradation of AXIN / Activation of NF-kappaB in B cells / Degradation of GLI1 by the proteasome / Hedgehog ligand biogenesis / P-body / G2/M Checkpoints / Defective CFTR causes cystic fibrosis / Negative regulation of NOTCH4 signaling / GSK3B and BTRC:CUL1-mediated-degradation of NFE2L2 / Autodegradation of the E3 ubiquitin ligase COP1 / Vif-mediated degradation of APOBEC3G Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.47 Å | |||||||||
Authors | Arkinson C / Gee CL / Martin A | |||||||||
Funding support | United States, 1 items
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Citation | Journal: bioRxiv / Year: 2024 Title: Structural landscape of AAA+ ATPase motor states in the substrate-degrading human 26S proteasome reveals conformation-specific binding of TXNL1. Authors: Connor Arkinson / Christine L Gee / Zeyuan Zhang / Ken C Dong / Andreas Martin / Abstract: The 26S proteasome targets many cellular proteins for degradation during general homeostasis, protein quality control, and the regulation of vital processes. A broad range of proteasome-interacting ...The 26S proteasome targets many cellular proteins for degradation during general homeostasis, protein quality control, and the regulation of vital processes. A broad range of proteasome-interacting cofactors thereby modulates these functions and aids in substrate degradation. Here, we solved several high-resolution structures of the redox active cofactor TXNL1 bound to the human 26S proteasome at saturating and sub-stoichiometric concentrations by time resolved cryo-EM. We identified distinct binding modes of TXNL1 that depend on the proteasome conformational and ATPase motor states. Together with biophysical and biochemical experiments, our structural studies reveal that the resting-state proteasome prior to substrate engagement with the ATPase motor binds TXNL1 with low affinity and in variable positions on top of the Rpn11 deubiquitinase. In contrast, the actively degrading proteasome shows additional interactions leading to high-affinity TXNL1 binding, whereby TXNL1's C-terminal tail covers the catalytic groove of the Rpn11 deubiquitinase and coordinates the active-site Zn. Furthermore, these cryo-EM structures of the degrading proteasome capture the ATPase hexamer in all registers of spiral-staircase arrangements and thus visualize the complete ATP-hydrolysis cycle of the AAA+ motor, indicating temporally asymmetric hydrolysis and conformational changes in bursts during mechanical substrate unfolding and translocation. Remarkably, we catch the proteasome in the act of unfolding the beta-barrel mEos3.2 substrate while the ATPase hexamer is in a particular spiral staircase register. Our findings challenge current models for protein translocation through hexameric AAA+ motors and reveal how the proteasome uses its distinct but broad range of conformational states to coordinate cofactor binding and substrate processing. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_47722.map.gz | 75.4 MB | EMDB map data format | |
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Header (meta data) | emd-47722-v30.xml emd-47722.xml | 51.3 KB 51.3 KB | Display Display | EMDB header |
Images | emd_47722.png | 56.3 KB | ||
Filedesc metadata | emd-47722.cif.gz | 13.8 KB | ||
Others | emd_47722_additional_1.map.gz emd_47722_half_map_1.map.gz emd_47722_half_map_2.map.gz | 134.7 MB 139 MB 139 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-47722 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-47722 | HTTPS FTP |
-Validation report
Summary document | emd_47722_validation.pdf.gz | 1.3 MB | Display | EMDB validaton report |
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Full document | emd_47722_full_validation.pdf.gz | 1.3 MB | Display | |
Data in XML | emd_47722_validation.xml.gz | 14.7 KB | Display | |
Data in CIF | emd_47722_validation.cif.gz | 17.3 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-47722 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-47722 | HTTPS FTP |
-Related structure data
Related structure data | 9e8jMC 9e8gC 9e8hC 9e8iC 9e8kC 9e8lC 9e8nC 9e8oC 9e8qC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_47722.map.gz / Format: CCP4 / Size: 149.9 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: 1.048 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Additional map: Sharpened
File | emd_47722_additional_1.map | ||||||||||||
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Annotation | Sharpened | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_47722_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_47722_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Human 26S proteasome complexed with Nub1, Fat10 and TXNL1
+Supramolecule #1: Human 26S proteasome complexed with Nub1, Fat10 and TXNL1
+Macromolecule #1: 26S proteasome regulatory subunit 7
+Macromolecule #2: 26S proteasome regulatory subunit 4
+Macromolecule #3: 26S protease regulatory subunit 8
+Macromolecule #4: 26S proteasome regulatory subunit 6B
+Macromolecule #5: 26S protease regulatory subunit 10B
+Macromolecule #6: 26S proteasome regulatory subunit 6A
+Macromolecule #7: Proteasome subunit alpha type-6
+Macromolecule #8: Proteasome subunit alpha type-2
+Macromolecule #9: Proteasome subunit alpha type-4
+Macromolecule #10: Proteasome subunit alpha type-7
+Macromolecule #11: Proteasome subunit alpha type-5
+Macromolecule #12: Proteasome subunit alpha type-1
+Macromolecule #13: Proteasome subunit alpha type-3
+Macromolecule #14: 26S proteasome non-ATPase regulatory subunit 1
+Macromolecule #15: 26S proteasome non-ATPase regulatory subunit 3
+Macromolecule #16: 26S proteasome non-ATPase regulatory subunit 12
+Macromolecule #17: 26S proteasome non-ATPase regulatory subunit 11
+Macromolecule #18: 26S proteasome non-ATPase regulatory subunit 6
+Macromolecule #19: 26S proteasome non-ATPase regulatory subunit 7
+Macromolecule #20: 26S proteasome non-ATPase regulatory subunit 13
+Macromolecule #21: 26S proteasome non-ATPase regulatory subunit 4
+Macromolecule #22: 26S proteasome non-ATPase regulatory subunit 14
+Macromolecule #23: 26S proteasome non-ATPase regulatory subunit 8
+Macromolecule #24: 26S proteasome complex subunit SEM1
+Macromolecule #25: 26S proteasome non-ATPase regulatory subunit 2
+Macromolecule #26: Isoform 2 of NEDD8 ultimate buster 1
+Macromolecule #27: Thioredoxin-like protein 1
+Macromolecule #28: substrate peptide
+Macromolecule #29: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #30: MAGNESIUM ION
+Macromolecule #31: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #32: ZINC ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.4 Details: 30 mM HEPES pH7.4, 25 mM NaCl, 25 mM KCl, 3% (v/v) glycerol, 5 mM MgCl2 2 mM ATP and 0.5 mM TCEP. |
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Grid | Model: UltrAuFoil R2/2 / Material: GOLD / Mesh: 200 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 25 sec. / Details: 25 mA |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 285 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 50.0 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: 1.7 µm / Nominal defocus min: 0.5 µm |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: NONE |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 3.47 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 19309 |
Initial angle assignment | Type: MAXIMUM LIKELIHOOD |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |