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Yorodumi- PDB-8jrt: Cryo-EM structure of human 26S proteasomal RP subcomplex (Ea stat... -
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Basic information
| Entry | Database: PDB / ID: 8jrt | |||||||||||||||
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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. | |||||||||||||||
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Keywords | CYTOSOLIC PROTEIN / protein degradation / macromolecular complex / ubiquitin-proteasome system | |||||||||||||||
| 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 / hypothalamus gonadotrophin-releasing hormone neuron development / metal-dependent deubiquitinase activity / protein K63-linked deubiquitination / negative regulation of programmed cell death / female meiosis I / Regulation of ornithine decarboxylase (ODC) / positive regulation of protein monoubiquitination / Proteasome assembly / Cross-presentation of soluble exogenous antigens (endosomes) / fat pad development / proteasome core complex / mitochondrion transport along microtubule / 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 / female gonad development / Impaired BRCA2 binding to RAD51 / seminiferous tubule development / proteasome binding / transcription factor binding / regulation of protein catabolic process / myofibril / 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 / positive regulation of RNA polymerase II transcription preinitiation complex assembly / polyubiquitin modification-dependent protein binding / immune system process / protein deubiquitination / NF-kappaB binding / endopeptidase activator activity / mRNA export from nucleus / proteasome assembly / proteasome core complex, alpha-subunit complex / energy homeostasis / SARS-CoV-1 targets host intracellular signalling and regulatory pathways / regulation of neuron apoptotic process / neuron projection morphogenesis / enzyme regulator activity / regulation of proteasomal protein catabolic process / ERAD pathway / Maturation of protein E / Maturation of protein E / inclusion body / 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 / Endosomal Sorting Complex Required For Transport (ESCRT) / Membrane binding and targetting of GAG proteins / Negative regulation of FLT3 / Regulation of TBK1, IKKε (IKBKE)-mediated activation of IRF3, IRF7 / 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 / Downregulation of ERBB4 signaling / Regulation of FZD by ubiquitination / APC-Cdc20 mediated degradation of Nek2A / p75NTR recruits signalling complexes / InlA-mediated entry of Listeria monocytogenes into host cells / TRAF6 mediated IRF7 activation in TLR7/8 or 9 signaling / TRAF6-mediated induction of TAK1 complex within TLR4 complex / Regulation of pyruvate metabolism / NF-kB is activated and signals survival / Regulation of innate immune responses to cytosolic DNA Similarity search - Function | |||||||||||||||
| Biological species | Homo sapiens (human) | |||||||||||||||
| Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.6 Å | |||||||||||||||
Authors | Hsu, S.T.D. / Draczkowski, P. / Wang, Y.S. | |||||||||||||||
| Funding support | Taiwan, 4items
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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
| Structure viewer | Molecule: Molmil Jmol/JSmol |
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Downloads & links
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Download
| PDBx/mmCIF format | 8jrt.cif.gz | 1.5 MB | Display | PDBx/mmCIF format |
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| PDB format | pdb8jrt.ent.gz | Display | PDB format | |
| PDBx/mmJSON format | 8jrt.json.gz | Tree view | PDBx/mmJSON format | |
| Others | Other downloads |
-Validation report
| Summary document | 8jrt_validation.pdf.gz | 1.5 MB | Display | wwPDB validaton report |
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| Full document | 8jrt_full_validation.pdf.gz | 1.6 MB | Display | |
| Data in XML | 8jrt_validation.xml.gz | 214.8 KB | Display | |
| Data in CIF | 8jrt_validation.cif.gz | 344.1 KB | Display | |
| Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/jr/8jrt ftp://data.pdbj.org/pub/pdb/validation_reports/jr/8jrt | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 36605MC ![]() 8jriC ![]() 8jtiC ![]() 8k0gC M: map data used to model this data C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
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Assembly
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Components
-26S protease regulatory subunit ... , 6 types, 6 molecules ABCDEF
| #1: Protein | Mass: 48700.805 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMC2, MSS1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P35998 |
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| #2: Protein | Mass: 49260.504 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMC1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P62191 |
| #3: Protein | Mass: 45694.047 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMC5, SUG1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P62195 |
| #4: Protein | Mass: 47426.141 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMC4, MIP224, TBP7 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P43686 |
| #19: Protein | Mass: 44241.008 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMC6, SUG2 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P62333 |
| #20: Protein | Mass: 49266.457 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMC3, TBP1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P17980 |
-Proteasome subunit alpha type- ... , 7 types, 7 molecules GHIJKLM
| #5: Protein | Mass: 27432.459 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMA6, PROS27 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human)References: UniProt: P60900, proteasome endopeptidase complex |
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| #6: Protein | Mass: 25927.535 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMA2, HC3, PSC3 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human)References: UniProt: P25787, proteasome endopeptidase complex |
| #7: Protein | Mass: 29525.842 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMA4, HC9, PSC9 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human)References: UniProt: P25789, proteasome endopeptidase complex |
| #8: Protein | Mass: 27929.891 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMA7, HSPC / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human)References: UniProt: O14818, proteasome endopeptidase complex |
| #9: Protein | Mass: 26435.977 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMA5 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human)References: UniProt: P28066, proteasome endopeptidase complex |
| #10: Protein | Mass: 29595.627 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMA1, HC2, NU, PROS30, PSC2 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human)References: UniProt: P25786, proteasome endopeptidase complex |
| #11: Protein | Mass: 28469.252 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMA3, HC8, PSC8 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P25788 |
-26S proteasome non-ATPase regulatory subunit ... , 11 types, 11 molecules VWXYdfUZabc
| #12: Protein | Mass: 61066.500 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD3 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: O43242 |
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| #13: Protein | Mass: 52979.359 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD12 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: O00232 |
| #14: Protein | Mass: 47526.688 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD11 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: O00231 |
| #15: Protein | Mass: 45592.285 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD6, KIAA0107, PFAAP4 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: Q15008 |
| #16: Protein | Mass: 39667.871 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD8 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P48556 |
| #18: Protein | Mass: 100313.625 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD2, TRAP2 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: Q13200 |
| #21: Protein | Mass: 105958.234 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: Q99460 |
| #22: Protein | Mass: 37086.441 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD7, MOV34L / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P51665 |
| #23: Protein | Mass: 42995.359 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD13 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: Q9UNM6 |
| #24: Protein | Mass: 40781.590 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD4, MCB1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P55036 |
| #25: Protein | Mass: 34620.023 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: PSMD14, POH1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human)References: UniProt: O00487, Hydrolases; Acting on peptide bonds (peptidases); Omega peptidases |
-Protein , 2 types, 4 molecules evuw
| #17: Protein | Mass: 8284.611 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Homo sapiens (human) / Gene: SHFM1, DSS1, SHFDG1 / Cell line (production host): HEK293 / Organ (production host): Kidney / Production host: Homo sapiens (human) / References: UniProt: P60896 |
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| #26: Protein | Mass: 8950.154 Da / Num. of mol.: 3 / Mutation: K63R Source method: isolated from a genetically manipulated source Details: GSGGS linker sequence remaining at C-terminus after TEV cleavage Source: (gene. exp.) Homo sapiens (human) / Gene: UBB / Production host: ![]() |
-Non-polymers , 3 types, 11 molecules 




| #27: Chemical | ChemComp-ATP / #28: Chemical | ChemComp-MG / #29: Chemical | |
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-Details
| Has ligand of interest | Y |
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| Has protein modification | Y |
-Experimental details
-Experiment
| Experiment | Method: ELECTRON MICROSCOPY |
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| EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
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Sample preparation
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| Molecular weight |
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| Buffer solution | pH: 7.6 | ||||||||||||||||||||||||||||||||||||||||||
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| Specimen | Conc.: 1 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES Details: The complex was additionally supplemented with an excess of preformed and SEC-purified RPN13:UCHL5 complex. | ||||||||||||||||||||||||||||||||||||||||||
| Specimen support | Details: 20 mA / Grid material: COPPER / Grid mesh size: 300 divisions/in. / Grid type: Quantifoil R1.2/1.3 | ||||||||||||||||||||||||||||||||||||||||||
| Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277.15 K / Details: incubation time= 3 s blotting time= 2.5 s |
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Electron microscopy imaging
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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| Microscopy | Model: FEI TITAN KRIOS |
| Electron gun | Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM |
| Electron lens | Mode: BRIGHT FIELD / Nominal magnification: 70000 X / Nominal defocus max: 1800 nm / Nominal defocus min: 1200 nm / Alignment procedure: COMA FREE |
| Specimen holder | Cryogen: NITROGEN / Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER |
| Image recording | Average exposure time: 1.8 sec. / Electron dose: 49 e/Å2 / Film or detector model: GATAN K3 (6k x 4k) / Num. of grids imaged: 3 / Num. of real images: 15242 |
| EM imaging optics | Energyfilter slit width: 20 eV |
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Processing
| EM software |
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| CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Particle selection | Num. of particles selected: 2695911 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Symmetry | Point symmetry: C1 (asymmetric) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| 3D reconstruction | Resolution: 3.6 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 54794 / Algorithm: FOURIER SPACE Details: Performed in cryoSPARC using Non-uniform Refinment and Local Refinment job types. Num. of class averages: 1 / Symmetry type: POINT | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | B value: 121.58 / Protocol: OTHER / Space: REAL / Target criteria: 0.83 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Atomic model building | PDB-ID: 6MSB Accession code: 6MSB / Source name: PDB / Type: experimental model | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Refine LS restraints |
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Homo sapiens (human)
Taiwan, 4items
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FIELD EMISSION GUN
