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Yorodumi- EMDB-1140: Localization of the coactivator Cdh1 and the cullin subunit Apc2 ... -
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Basic information
| Entry | Database: EMDB / ID: EMD-1140 | |||||||||
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| Title | Localization of the coactivator Cdh1 and the cullin subunit Apc2 in a cryo-electron microscopy model of vertebrate APC/C. | |||||||||
Map data | This is the 3D map of the Xenopus anaphase promoting complex (APC/C) | |||||||||
Sample |
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| Method | single particle reconstruction / cryo EM / negative staining / Resolution: 24.0 Å | |||||||||
Authors | Dube P / Herzog F / Peters JM / Stark H | |||||||||
Citation | Journal: Mol Cell / Year: 2005Title: Localization of the coactivator Cdh1 and the cullin subunit Apc2 in a cryo-electron microscopy model of vertebrate APC/C. Authors: Prakash Dube / Franz Herzog / Christian Gieffers / Bjoern Sander / Dietmar Riedel / Shirley A Müller / Andreas Engel / Jan-Michael Peters / Holger Stark / ![]() Abstract: The anaphase-promoting complex/cyclosome (APC/C) is a ubiquitin ligase with essential functions in mitosis, meiosis, and G1 phase of the cell cycle. APC/C recognizes substrates via coactivator ...The anaphase-promoting complex/cyclosome (APC/C) is a ubiquitin ligase with essential functions in mitosis, meiosis, and G1 phase of the cell cycle. APC/C recognizes substrates via coactivator proteins such as Cdh1, and bound substrates are ubiquitinated by E2 enzymes that interact with a hetero-dimer of the RING subunit Apc11 and the cullin Apc2. We have obtained three-dimensional (3D) models of human and Xenopus APC/C by angular reconstitution and random conical tilt (RCT) analyses of negatively stained cryo-electron microscopy (cryo-EM) preparations, have determined the masses of these particles by scanning transmission electron microscopy (STEM), and have mapped the locations of Cdh1 and Apc2. These proteins are located on the same side of the asymmetric APC/C, implying that this is where substrates are ubiquitinated. We have further identified a large flexible domain in APC/C that adopts a different orientation upon Cdh1 binding. Cdh1 may thus activate APC/C both by recruiting substrates and by inducing conformational changes. | |||||||||
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Structure visualization
| Movie |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
| Supplemental images |
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Downloads & links
-EMDB archive
| Map data | emd_1140.map.gz | 14.5 MB | EMDB map data format | |
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| Header (meta data) | emd-1140-v30.xml emd-1140.xml | 7.6 KB 7.6 KB | Display Display | EMDB header |
| Images | 1140.gif | 7.2 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-1140 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-1140 | HTTPS FTP |
-Validation report
| Summary document | emd_1140_validation.pdf.gz | 246.3 KB | Display | EMDB validaton report |
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| Full document | emd_1140_full_validation.pdf.gz | 245.5 KB | Display | |
| Data in XML | emd_1140_validation.xml.gz | 5.8 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-1140 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-1140 | HTTPS FTP |
-Related structure data
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Map
| File | Download / File: emd_1140.map.gz / Format: CCP4 / Size: 15.3 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | This is the 3D map of the Xenopus anaphase promoting complex (APC/C) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 3.318 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
CCP4 map header:
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-Supplemental data
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Sample components
-Entire : Anaphase Promoting Complex
| Entire | Name: Anaphase Promoting Complex |
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| Components |
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-Supramolecule #1000: Anaphase Promoting Complex
| Supramolecule | Name: Anaphase Promoting Complex / type: sample / ID: 1000 / Oligomeric state: Monomer / Number unique components: 1 |
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| Molecular weight | Experimental: 1.5 MDa / Method: STEM |
-Macromolecule #1: Anaphase Promoting Complex
| Macromolecule | Name: Anaphase Promoting Complex / type: protein_or_peptide / ID: 1 / Recombinant expression: No |
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| Source (natural) | Organism: |
-Experimental details
-Structure determination
| Method | negative staining, cryo EM |
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Processing | single particle reconstruction |
| Aggregation state | particle |
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Sample preparation
| Buffer | pH: 7.6 / Details: 20 mM Hepes, 150 mM NaCl, 0.5 mM DTT |
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| Staining | Type: NEGATIVE / Details: carbon sandwich in 2% w/v uranyl formate |
| Vitrification | Cryogen name: NITROGEN |
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Electron microscopy
| Microscope | FEI/PHILIPS CM200FEG |
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| Image recording | Category: CCD / Film or detector model: GENERIC TVIPS (4k x 4k) / Average electron dose: 15 e/Å2 |
| Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Calibrated magnification: 90500 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2 mm / Nominal magnification: 90500 |
| Sample stage | Specimen holder: Eucentric / Specimen holder model: GATAN LIQUID NITROGEN |
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Image processing
| CTF correction | Details: Each particle |
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| Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 24.0 Å / Resolution method: OTHER / Software - Name: Imagic-5 |
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