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Yorodumi- EMDB-21120: Ankyrin repeat and SOCS-box protein 9 (ASB9), ElonginB (ELOB), an... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-21120 | |||||||||
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Title | Ankyrin repeat and SOCS-box protein 9 (ASB9), ElonginB (ELOB), and ElonginC (ELOC) bound to its substrate Brain-type Creatine Kinase (CKB) | |||||||||
Map data | sharpened map | |||||||||
Sample |
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Keywords | Ankyrin repeat / Elongation Factor / Creatine Kinase / Ubiquitin / TRANSCRIPTION | |||||||||
Function / homology | Function and homology information futile creatine cycle / Creatine metabolism / creatine kinase / phosphocreatine biosynthetic process / creatine kinase activity / target-directed miRNA degradation / elongin complex / VCB complex / Cul5-RING ubiquitin ligase complex / RND3 GTPase cycle ...futile creatine cycle / Creatine metabolism / creatine kinase / phosphocreatine biosynthetic process / creatine kinase activity / target-directed miRNA degradation / elongin complex / VCB complex / Cul5-RING ubiquitin ligase complex / RND3 GTPase cycle / Cul2-RING ubiquitin ligase complex / Pausing and recovery of Tat-mediated HIV elongation / Tat-mediated HIV elongation arrest and recovery / HIV elongation arrest and recovery / Pausing and recovery of HIV elongation / ubiquitin-like ligase-substrate adaptor activity / Tat-mediated elongation of the HIV-1 transcript / Formation of HIV-1 elongation complex containing HIV-1 Tat / Formation of HIV elongation complex in the absence of HIV Tat / RNA Polymerase II Transcription Elongation / Formation of RNA Pol II elongation complex / RNA Polymerase II Pre-transcription Events / substantia nigra development / transcription corepressor binding / TP53 Regulates Transcription of DNA Repair Genes / transcription initiation at RNA polymerase II promoter / transcription elongation by RNA polymerase II / Vif-mediated degradation of APOBEC3G / Inactivation of CSF3 (G-CSF) signaling / Oxygen-dependent proline hydroxylation of Hypoxia-inducible Factor Alpha / Evasion by RSV of host interferon responses / Regulation of expression of SLITs and ROBOs / positive regulation of protein catabolic process / Antigen processing: Ubiquitination & Proteasome degradation / positive regulation of proteasomal ubiquitin-dependent protein catabolic process / Neddylation / protein-macromolecule adaptor activity / ubiquitin-dependent protein catabolic process / protein-containing complex assembly / proteasome-mediated ubiquitin-dependent protein catabolic process / intracellular signal transduction / protein ubiquitination / ubiquitin protein ligase binding / regulation of transcription by RNA polymerase II / mitochondrion / extracellular space / extracellular exosome / nucleoplasm / ATP binding / plasma membrane / cytosol Similarity search - Function | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.1 Å | |||||||||
Authors | Komives EA / Lumpkin RJ | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Nat Commun / Year: 2020 Title: Structure and dynamics of the ASB9 CUL-RING E3 Ligase. Authors: Ryan J Lumpkin / Richard W Baker / Andres E Leschziner / Elizabeth A Komives / Abstract: The Cullin 5 (CUL5) Ring E3 ligase uses adaptors Elongins B and C (ELOB/C) to bind different SOCS-box-containing substrate receptors, determining the substrate specificity of the ligase. The 18- ...The Cullin 5 (CUL5) Ring E3 ligase uses adaptors Elongins B and C (ELOB/C) to bind different SOCS-box-containing substrate receptors, determining the substrate specificity of the ligase. The 18-member ankyrin and SOCS box (ASB) family is the largest substrate receptor family. Here we report cryo-EM data for the substrate, creatine kinase (CKB) bound to ASB9-ELOB/C, and for full-length CUL5 bound to the RING protein, RBX2, which binds various E2s. To date, no full structures are available either for a substrate-bound ASB nor for CUL5. Hydrogen-deuterium exchange (HDX-MS) mapped onto a full structural model of the ligase revealed long-range allostery extending from the substrate through CUL5. We propose a revised allosteric mechanism for how CUL-E3 ligases function. ASB9 and CUL5 behave as rigid rods, connected through a hinge provided by ELOB/C transmitting long-range allosteric crosstalk from the substrate through CUL5 to the RBX2 flexible linker. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_21120.map.gz | 97 MB | EMDB map data format | |
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Header (meta data) | emd-21120-v30.xml emd-21120.xml | 22.4 KB 22.4 KB | Display Display | EMDB header |
Images | emd_21120.png | 131.5 KB | ||
Filedesc metadata | emd-21120.cif.gz | 6.3 KB | ||
Others | emd_21120_additional.map.gz emd_21120_half_map_1.map.gz emd_21120_half_map_2.map.gz | 51.3 MB 95.5 MB 95.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21120 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21120 | HTTPS FTP |
-Validation report
Summary document | emd_21120_validation.pdf.gz | 821 KB | Display | EMDB validaton report |
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Full document | emd_21120_full_validation.pdf.gz | 820.6 KB | Display | |
Data in XML | emd_21120_validation.xml.gz | 13.4 KB | Display | |
Data in CIF | emd_21120_validation.cif.gz | 15.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21120 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21120 | HTTPS FTP |
-Related structure data
Related structure data | 6v9hMC 6v9iC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_21120.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | sharpened map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.16 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Additional map: unfiltered map
File | emd_21120_additional.map | ||||||||||||
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Annotation | unfiltered map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map 1
File | emd_21120_half_map_1.map | ||||||||||||
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Annotation | half map 1 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: half map 2
File | emd_21120_half_map_2.map | ||||||||||||
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Annotation | half map 2 | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Ankyrin repeat and SOCS-box protein 9 (ASB9), ElonginB (ELOB), an...
Entire | Name: Ankyrin repeat and SOCS-box protein 9 (ASB9), ElonginB (ELOB), and ElonginC (ELOC) bound to its substrate Brain-type Creatine Kinase (CKB) |
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Components |
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-Supramolecule #1: Ankyrin repeat and SOCS-box protein 9 (ASB9), ElonginB (ELOB), an...
Supramolecule | Name: Ankyrin repeat and SOCS-box protein 9 (ASB9), ElonginB (ELOB), and ElonginC (ELOC) bound to its substrate Brain-type Creatine Kinase (CKB) type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 140 KDa |
-Macromolecule #1: Creatine kinase B-type
Macromolecule | Name: Creatine kinase B-type / type: protein_or_peptide / ID: 1 / Number of copies: 2 / Enantiomer: LEVO / EC number: creatine kinase |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 42.699207 KDa |
Recombinant expression | Organism: Escherichia coli BL21 (bacteria) |
Sequence | String: MPFSNSHNAL KLRFPAEDEF PDLSAHNNHM AKVLTPELYA ELRAKSTPSG FTLDDVIQTG VDNPGHPYIM TVGCVAGDEE SYEVFKDLF DPIIEDRHGG YKPSDEHKTD LNPDNLQGGD DLDPNYVLSS RVRTGRSIRG FCLPPHCSRG ERRAIEKLAV E ALSSLDGD ...String: MPFSNSHNAL KLRFPAEDEF PDLSAHNNHM AKVLTPELYA ELRAKSTPSG FTLDDVIQTG VDNPGHPYIM TVGCVAGDEE SYEVFKDLF DPIIEDRHGG YKPSDEHKTD LNPDNLQGGD DLDPNYVLSS RVRTGRSIRG FCLPPHCSRG ERRAIEKLAV E ALSSLDGD LAGRYYALKS MTEAEQQQLI DDHFLFDKPV SPLLLASGMA RDWPDARGIW HNDNKTFLVW VNEEDHLRVI SM QKGGNMK EVFTRFCTGL TQIETLFKSK DYEFMWNPHL GYILTCPSNL GTGLRAGVHI KLPNLGKHEK FSEVLKRLRL QKR GTGGVD TAAVGGVFDV SNADRLGFSE VELVQMVVDG VKLLIEMEQR LEQGQAIDDL MPAQK UniProtKB: Creatine kinase B-type |
-Macromolecule #2: Ankyrin repeat and SOCS box protein 9
Macromolecule | Name: Ankyrin repeat and SOCS box protein 9 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 34.186141 KDa |
Recombinant expression | Organism: Escherichia coli BL21 (bacteria) |
Sequence | String: MKHHHHHHHH GGLVPRGSHG MDGKQGGMDG SKPAGPRDFP GIRLLSNPLM GDAVSDWSPM HEAAIHGHQL SLRNLISQGW AVNIITADH VSPLHEACLG GHLSCVKILL KHGAQVNGVT ADWHTPLFNA CVSGSWDCVN LLLQHGASVQ PESDLASPIH E AARRGHVE ...String: MKHHHHHHHH GGLVPRGSHG MDGKQGGMDG SKPAGPRDFP GIRLLSNPLM GDAVSDWSPM HEAAIHGHQL SLRNLISQGW AVNIITADH VSPLHEACLG GHLSCVKILL KHGAQVNGVT ADWHTPLFNA CVSGSWDCVN LLLQHGASVQ PESDLASPIH E AARRGHVE CVNSLIAYGG NIDHKISHLG TPLYLACENQ QRACVKKLLE SGADVNQGKG QDSPLHAVAR TASEELACLL MD FGADTQA KNAEGKRPVE LVPPESPLAQ LFLEREGPPS LMQLCRLRIR KCFGIQQHHK ITKLVLPEDL KQFLLHL UniProtKB: Ankyrin repeat and SOCS box protein 9 |
-Macromolecule #3: Elongin-C
Macromolecule | Name: Elongin-C / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 10.974616 KDa |
Recombinant expression | Organism: Escherichia coli BL21 (bacteria) |
Sequence | String: MMYVKLISSD GHEFIVKREH ALTSGTIKAM LSGPGQFAEN ETNEVNFREI PSHVLSKVCM YFTYKVRYTN SSTEIPEFPI APEIALELL MAANFLDC UniProtKB: Elongin-C |
-Macromolecule #4: Elongin-B
Macromolecule | Name: Elongin-B / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 13.147781 KDa |
Recombinant expression | Organism: Escherichia coli BL21 (bacteria) |
Sequence | String: MDVFLMIRRH KTTIFTDAKE SSTVFELKRI VEGILKRPPD EQRLYKDDQL LDDGKTLGEC GFTSQTARPQ APATVGLAFR ADDTFEALC IEPFSSPPEL PDVMKPQDSG SSANEQAVQ UniProtKB: Elongin-B |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 3 mg/mL |
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Buffer | pH: 7.5 |
Grid | Model: UltrAuFoil / Material: GOLD / Mesh: 300 / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 30 sec. / Pretreatment - Atmosphere: AIR |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: 4 second blot, blot force 20. |
-Electron microscopy
Microscope | FEI TALOS ARCTICA |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average exposure time: 7.0 sec. / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 70.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 2.5 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 36000 |
Sample stage | Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Talos Arctica / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Initial model |
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Refinement | Space: REAL / Protocol: FLEXIBLE FIT | ||||||
Output model | PDB-6v9h: |