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Yorodumi- EMDB-71740: Human 26S proteasome bound to TXNL1 with opened gate of core particle -
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Open data
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Basic information
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| Title | Human 26S proteasome bound to TXNL1 with opened gate of core particle | |||||||||
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Keywords | Proteasome / 26S / TXNL1 / HYDROLASE | |||||||||
| Function / homology | Function and homology informationdisulfide oxidoreductase activity / positive regulation of inclusion body assembly / thyrotropin-releasing hormone receptor binding / nuclear proteasome complex / host-mediated perturbation of viral transcription / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / proteasome accessory complex / purine ribonucleoside triphosphate binding / integrator complex ...disulfide oxidoreductase activity / positive regulation of inclusion body assembly / thyrotropin-releasing hormone receptor binding / nuclear proteasome complex / host-mediated perturbation of viral transcription / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / proteasome accessory complex / purine ribonucleoside triphosphate binding / integrator complex / proteasome regulatory particle / cytosolic proteasome complex / positive regulation of proteasomal protein catabolic process / CD8-positive, alpha-beta T cell differentiation / CD8-positive, alpha-beta T cell homeostasis / transcription factor binding / thymic T cell selection / Antigen processing: Ub, ATP-independent proteasomal degradation / proteasome-activating activity / RND1 GTPase cycle / RND2 GTPase cycle / RND3 GTPase cycle / proteasome regulatory particle, lid subcomplex / proteasome regulatory particle, base subcomplex / negative regulation of programmed cell death / RHOBTB1 GTPase cycle / negative regulation of regulatory T cell differentiation / T-helper 1 cell differentiation / protein K63-linked deubiquitination / RHOV GTPase cycle / metal-dependent deubiquitinase activity / Regulation of ornithine decarboxylase (ODC) / proteasome core complex / Proteasome assembly / cellular response to type I interferon / T-helper 17 cell differentiation / Cross-presentation of soluble exogenous antigens (endosomes) / retrograde vesicle-mediated transport, Golgi to endoplasmic reticulum / Somitogenesis / K63-linked deubiquitinase 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) / flagellated sperm motility / Resolution of D-loop Structures through Holliday Junction Intermediates / sperm end piece / proteasome binding / RHOU GTPase cycle / Impaired BRCA2 binding to RAD51 / myofibril / protein-disulfide reductase activity / ciliary tip / proteasomal ubiquitin-independent protein catabolic process / positive regulation of RNA polymerase II transcription preinitiation complex assembly / general transcription initiation factor binding / Presynaptic phase of homologous DNA pairing and strand exchange / proteasome storage granule / polyubiquitin modification-dependent protein binding / protein deubiquitination / NF-kappaB binding / RHOBTB2 GTPase cycle / endopeptidase activator activity / proteasome core complex, alpha-subunit complex / proteasome assembly / mRNA export from nucleus / SARS-CoV-1 targets host intracellular signalling and regulatory pathways / immune system process / regulation of G1/S transition of mitotic cell cycle / stem cell differentiation / regulation of macroautophagy / positive regulation of interleukin-2 production / proteasome complex / enzyme regulator activity / ERAD pathway / response to type II interferon / inclusion body / TBP-class protein binding / : / regulation of proteasomal protein catabolic process / sarcomere / Regulation of activated PAK-2p34 by proteasome mediated degradation / ubiquitin binding / negative regulation of inflammatory response to antigenic stimulus / Autodegradation of Cdh1 by Cdh1:APC/C / proteasomal protein catabolic process / 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 / lipopolysaccharide binding / SCF-beta-TrCP mediated degradation of Emi1 / NIK-->noncanonical NF-kB signaling / AUF1 (hnRNP D0) binds and destabilizes mRNA / TNFR2 non-canonical NF-kB pathway / positive regulation of type II interferon production / Assembly of the pre-replicative complex / P-body / Vpu mediated degradation of CD4 Similarity search - Function | |||||||||
| Biological species | Homo sapiens (human) | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.82 Å | |||||||||
Authors | Chen X / Negi H / Walters KJ | |||||||||
| Funding support | United States, 1 items
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Citation | Journal: Mol Cell / Year: 2026Title: Structures of dynamic interactors at native proteasomes by PhIX-MS and cryo-electron microscopy. Authors: Kitaik Lee / Hitendra Negi / Xiang Chen / Katerina Atallah-Yunes / Sophia Truslow / Rithik E Castelino / Mary R Guest / Anthony M Ciancone / Xiuxiu Lu / Sergey G Tarasov / Raj Chari / Kylie ...Authors: Kitaik Lee / Hitendra Negi / Xiang Chen / Katerina Atallah-Yunes / Sophia Truslow / Rithik E Castelino / Mary R Guest / Anthony M Ciancone / Xiuxiu Lu / Sergey G Tarasov / Raj Chari / Kylie J Walters / Francis J O'Reilly / ![]() Abstract: Molecular machines rely on dynamic, low-affinity interactions to perform their functional roles. We developed PhIX-MS (photo-induced in situ crosslinking-mass spectrometry), a structural proteomics ...Molecular machines rely on dynamic, low-affinity interactions to perform their functional roles. We developed PhIX-MS (photo-induced in situ crosslinking-mass spectrometry), a structural proteomics workflow to capture topological information for such transient interactions in cells by UV-activated crosslinking. Applying PhIX-MS with cryo-electron microscopy (cryo-EM) to proteasomes, we mapped the redox sensor TXNL1 at the proteasome regulatory particle (RP), including its dynamic thioredoxin-like domain near RPN2/PSMD1 and RPN13/ADRM1, where it is ideal for reducing substrates prior to proteolysis. RPs without the proteolytic core particle (CP) were structurally resolved while bound to TXNL1 and/or the chaperone PSMD5/S5b, which inserts its C terminus into the ATPase pore, causing extensive structural rearrangements. Additionally, PhIX-MS and AlphaFold identified the ubiquitin ligase UBE3C/Hul5 at RPN2, RPN3, and a dynamic RPN10 region, tethering UBE3C above the substrate entry channel. Our integrative approach enables the localization of native, low-affinity protein interactions and is broadly applicable to dynamic macromolecular assemblies. | |||||||||
| History |
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_71740.map.gz | 210.4 MB | EMDB map data format | |
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| Header (meta data) | emd-71740-v30.xml emd-71740.xml | 75.2 KB 75.2 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_71740_fsc.xml | 15.9 KB | Display | FSC data file |
| Images | emd_71740.png | 64.6 KB | ||
| Filedesc metadata | emd-71740.cif.gz | 15.6 KB | ||
| Others | emd_71740_additional_1.map.gz emd_71740_half_map_1.map.gz emd_71740_half_map_2.map.gz | 274.1 MB 391.2 MB 391.1 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-71740 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-71740 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 9pmoMC ![]() 12bmC ![]() 9pmjC ![]() 9pmqC ![]() 9proC ![]() 9prtC ![]() 71736 C: citing same article ( M: atomic model generated by this map |
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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_71740.map.gz / Format: CCP4 / Size: 421.9 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: 0.81 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Additional map: #1
| File | emd_71740_additional_1.map | ||||||||||||
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-Half map: #2
| File | emd_71740_half_map_1.map | ||||||||||||
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-Half map: #1
| File | emd_71740_half_map_2.map | ||||||||||||
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Sample components
+Entire : Human proteasome from HCT116 cells
+Supramolecule #1: Human proteasome from HCT116 cells
+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: Isoform Long of 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: Thioredoxin-like protein 1
+Macromolecule #27: PHOSPHOTHIOPHOSPHORIC ACID-ADENYLATE ESTER
+Macromolecule #28: MAGNESIUM ION
+Macromolecule #29: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #30: ZINC ION
-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 | 0.75 mg/mL | ||||||||||||||||||
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| Buffer | pH: 7.5 Component:
Details: 50 mM Tris [pH7.5], 50 mM NaCl, 1.5 mM ATP-gamma-S, 5 mM MgCl2 and 2 mM DTT | ||||||||||||||||||
| Grid | Model: Quantifoil R1.2/1.3 / Material: COPPER / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR / Details: 25 mA | ||||||||||||||||||
| Vitrification | Cryogen name: ETHANE / Chamber humidity: 85 % / Chamber temperature: 295.15 K / Instrument: LEICA EM GP | ||||||||||||||||||
| Details | Quantifoil copper R 1.2/1.3 holey carbon 300 mesh grids (#Q3100CR1.3; Electron Microscopy Sciences). |
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Electron microscopy
| Microscope | FEI TALOS ARCTICA |
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| Specialist optics | Energy filter - Name: GIF Bioquantum |
| Image recording | Film or detector model: GATAN K3 (6k x 4k) / Digitization - Dimensions - Width: 5760 pixel / Digitization - Dimensions - Height: 4092 pixel / Number real images: 40 / Average exposure time: 2.5 sec. / Average electron dose: 55.6 e/Å2 |
| Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.8 µm / Nominal magnification: 100000 |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
| Initial model |
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| Refinement | Space: REAL / Protocol: RIGID BODY FIT / Overall B value: 220 / Target criteria: Cross-correlation coefficient | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Output model | ![]() PDB-9pmo: |
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About Yorodumi



Keywords
Homo sapiens (human)
Authors
United States, 1 items
Citation































Z (Sec.)
Y (Row.)
X (Col.)














































FIELD EMISSION GUN




