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- EMDB-36605: Cryo-EM structure of human 26S proteasomal RP subcomplex (Ea stat... -
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Basic information
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Title | Cryo-EM structure of human 26S proteasomal RP subcomplex (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub. | |||||||||||||||
![]() | Composite sharpened cryo-EM half map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub. | |||||||||||||||
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![]() | protein degradation / macromolecular complex / ubiquitin-proteasome system / CYTOSOLIC PROTEIN | |||||||||||||||
Function / homology | ![]() positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / integrator complex / proteasome accessory complex / purine ribonucleoside triphosphate binding / meiosis I ...positive regulation of inclusion body assembly / Impaired BRCA2 translocation to the nucleus / Impaired BRCA2 binding to SEM1 (DSS1) / thyrotropin-releasing hormone receptor binding / modulation by host of viral transcription / Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases / integrator complex / proteasome accessory complex / purine ribonucleoside triphosphate binding / meiosis I / cytosolic proteasome complex / proteasome regulatory particle / positive regulation of proteasomal protein catabolic process / proteasome regulatory particle, lid subcomplex / proteasome-activating activity / proteasome regulatory particle, base subcomplex / metal-dependent deubiquitinase activity / hypothalamus gonadotrophin-releasing hormone neuron development / negative regulation of programmed cell death / protein K63-linked deubiquitination / Regulation of ornithine decarboxylase (ODC) / Proteasome assembly / 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) / female meiosis I / Cross-presentation of soluble exogenous antigens (endosomes) / positive regulation of protein monoubiquitination / Resolution of D-loop Structures through Holliday Junction Intermediates / proteasome core complex / Somitogenesis / fat pad development / mitochondrion transport along microtubule / K63-linked deubiquitinase activity / Impaired BRCA2 binding to RAD51 / proteasome binding / female gonad development / myofibril / seminiferous tubule development / transcription factor binding / regulation of protein catabolic process / immune system process / male meiosis I / proteasome storage granule / Presynaptic phase of homologous DNA pairing and strand exchange / positive regulation of intrinsic apoptotic signaling pathway by p53 class mediator / general transcription initiation factor binding / blastocyst development / polyubiquitin modification-dependent protein binding / positive regulation of RNA polymerase II transcription preinitiation complex assembly / protein deubiquitination / endopeptidase activator activity / NF-kappaB binding / proteasome assembly / proteasome core complex, alpha-subunit complex / mRNA export from nucleus / SARS-CoV-1 targets host intracellular signalling and regulatory pathways / energy homeostasis / inclusion body / enzyme regulator activity / regulation of neuron apoptotic process / ERAD pathway / regulation of proteasomal protein catabolic process / Maturation of protein E / Maturation of protein E / ER Quality Control Compartment (ERQC) / Myoclonic epilepsy of Lafora / FLT3 signaling by CBL mutants / Prevention of phagosomal-lysosomal fusion / IRAK2 mediated activation of TAK1 complex / Alpha-protein kinase 1 signaling pathway / Glycogen synthesis / IRAK1 recruits IKK complex / IRAK1 recruits IKK complex upon TLR7/8 or 9 stimulation / Membrane binding and targetting of GAG proteins / Endosomal Sorting Complex Required For Transport (ESCRT) / Regulation of TBK1, IKKε (IKBKE)-mediated activation of IRF3, IRF7 / Negative regulation of FLT3 / PTK6 Regulates RTKs and Their Effectors AKT1 and DOK1 / Regulation of TBK1, IKKε-mediated activation of IRF3, IRF7 upon TLR3 ligation / Constitutive Signaling by NOTCH1 HD Domain Mutants / IRAK2 mediated activation of TAK1 complex upon TLR7/8 or 9 stimulation / NOTCH2 Activation and Transmission of Signal to the Nucleus / TICAM1,TRAF6-dependent induction of TAK1 complex / TICAM1-dependent activation of IRF3/IRF7 / APC/C:Cdc20 mediated degradation of Cyclin B / Regulation of FZD by ubiquitination / Downregulation of ERBB4 signaling / p75NTR recruits signalling complexes / APC-Cdc20 mediated degradation of Nek2A / TRAF6 mediated IRF7 activation in TLR7/8 or 9 signaling / InlA-mediated entry of Listeria monocytogenes into host cells / Regulation of innate immune responses to cytosolic DNA / TRAF6-mediated induction of TAK1 complex within TLR4 complex / Regulation of pyruvate metabolism / proteasome complex / NF-kB is activated and signals survival / Downregulation of ERBB2:ERBB3 signaling Similarity search - Function | |||||||||||||||
Biological species | ![]() | |||||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||||||||
![]() | Hsu STD / Draczkowski P / Wang YS | |||||||||||||||
Funding support | ![]()
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![]() | ![]() Title: Cryo-EM structure of human 26S proteasomal RP subcomplex bound with branched Ub chain. Authors: Draczkowski P / Hsu STD / Wang YS | |||||||||||||||
History |
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Structure visualization
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 220 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 61.6 KB 61.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.2 KB | Display | ![]() |
Images | ![]() | 165.7 KB | ||
Filedesc metadata | ![]() | 14.6 KB | ||
Others | ![]() | 122.9 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 604.9 KB | Display | ![]() |
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Full document | ![]() | 604.4 KB | Display | |
Data in XML | ![]() | 13.8 KB | Display | |
Data in CIF | ![]() | 18.8 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8jrtMC ![]() 8jriC ![]() 8jtiC ![]() 8k0gC C: citing same article ( M: atomic model generated by this map |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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Annotation | Composite sharpened cryo-EM half map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub. | ||||||||||||||||||||||||||||||||||||
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: Composite unsharpened cryo-EM half map of human 26S...
File | emd_36605_additional_1.map | ||||||||||||
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Annotation | Composite unsharpened cryo-EM half map of human 26S RP (Ea state) bound to K11/K48-branched ubiquitin (Ub) chain composed of three Ub. | ||||||||||||
Projections & Slices |
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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...
+Supramolecule #5: K11/K48-branched ubiquitin (Ub) chain composed of three Ub
+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 3
+Macromolecule #13: 26S proteasome non-ATPase regulatory subunit 12
+Macromolecule #14: 26S proteasome non-ATPase regulatory subunit 11
+Macromolecule #15: 26S proteasome non-ATPase regulatory subunit 6
+Macromolecule #16: 26S proteasome non-ATPase regulatory subunit 8
+Macromolecule #17: 26S proteasome complex subunit DSS1
+Macromolecule #18: 26S proteasome non-ATPase regulatory subunit 2
+Macromolecule #19: 26S protease regulatory subunit 10B
+Macromolecule #20: 26S protease regulatory subunit 6A
+Macromolecule #21: 26S proteasome non-ATPase regulatory subunit 1
+Macromolecule #22: 26S proteasome non-ATPase regulatory subunit 7
+Macromolecule #23: 26S proteasome non-ATPase regulatory subunit 13
+Macromolecule #24: 26S proteasome non-ATPase regulatory subunit 4
+Macromolecule #25: 26S proteasome non-ATPase regulatory subunit 14
+Macromolecule #26: Polyubiquitin-B
+Macromolecule #27: ADENOSINE-5'-TRIPHOSPHATE
+Macromolecule #28: MAGNESIUM ION
+Macromolecule #29: ADENOSINE-5'-DIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | 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 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: ![]() |
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: 121.58 / Target criteria: 0.83 |
Output model | ![]() PDB-8jrt: |