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Yorodumi- EMDB-14099: STRUCTURE OF THE HUMAN INNER KINETOCHORE CCAN COMPLEX AT 10A RESO... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-14099 | |||||||||
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Title | STRUCTURE OF THE HUMAN INNER KINETOCHORE CCAN COMPLEX AT 10A RESOLUTION | |||||||||
Map data | main map | |||||||||
Sample |
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Keywords | INNER KINETOCHORE / CCAN / COMPLEX / DNA BINDING PROTEIN / CELL CYCLE | |||||||||
Biological species | Homo sapiens (human) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 10.0 Å | |||||||||
Authors | Vetter IR / Pesenti M / Raisch T | |||||||||
Funding support | 1 items
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Citation | Journal: Mol Cell / Year: 2022 Title: Structure of the human inner kinetochore CCAN complex and its significance for human centromere organization. Authors: Marion E Pesenti / Tobias Raisch / Duccio Conti / Kai Walstein / Ingrid Hoffmann / Dorothee Vogt / Daniel Prumbaum / Ingrid R Vetter / Stefan Raunser / Andrea Musacchio / Abstract: Centromeres are specialized chromosome loci that seed the kinetochore, a large protein complex that effects chromosome segregation. A 16-subunit complex, the constitutive centromere associated ...Centromeres are specialized chromosome loci that seed the kinetochore, a large protein complex that effects chromosome segregation. A 16-subunit complex, the constitutive centromere associated network (CCAN), connects between the specialized centromeric chromatin, marked by the histone H3 variant CENP-A, and the spindle-binding moiety of the kinetochore. Here, we report a cryo-electron microscopy structure of human CCAN. We highlight unique features such as the pseudo GTPase CENP-M and report how a crucial CENP-C motif binds the CENP-LN complex. The CCAN structure has implications for the mechanism of specific recognition of the CENP-A nucleosome. A model consistent with our structure depicts the CENP-C-bound nucleosome as connected to the CCAN through extended, flexible regions of CENP-C. An alternative model identifies both CENP-C and CENP-N as specificity determinants but requires CENP-N to bind CENP-A in a mode distinct from the classical nucleosome octamer. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_14099.map.gz | 7.1 MB | EMDB map data format | |
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Header (meta data) | emd-14099-v30.xml emd-14099.xml | 13.1 KB 13.1 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_14099_fsc.xml emd_14099_fsc_2.xml emd_14099_fsc_3.xml | 4.7 KB 4.7 KB 4.7 KB | Display Display Display | FSC data file |
Images | emd_14099.png | 159.9 KB | ||
Masks | emd_14099_msk_1.map emd_14099_msk_2.map emd_14099_msk_3.map | 8 MB 8 MB 8 MB | Mask map | |
Filedesc metadata | emd-14099.cif.gz | 4.6 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-14099 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-14099 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_14099.map.gz / Format: CCP4 / Size: 8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | main map | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 2.72 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_14099_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Mask #2
File | emd_14099_msk_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Mask #3
File | emd_14099_msk_3.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : human inner kinetochore CCAN complex
Entire | Name: human inner kinetochore CCAN complex |
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Components |
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-Supramolecule #1: human inner kinetochore CCAN complex
Supramolecule | Name: human inner kinetochore CCAN complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#15 |
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Source (natural) | Organism: Homo sapiens (human) |
Molecular weight | Theoretical: 451 KDa |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 1.5 mg/mL |
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Buffer | pH: 6.8 / Details: 0.0025% Triton |
Grid | Model: UltrAuFoil R1.2/1.3 / Material: GOLD / Pretreatment - Type: GLOW DISCHARGE |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 286 K / Instrument: FEI VITROBOT MARK IV |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 1.2 µm / Nominal defocus min: 0.6 µm / Nominal magnification: 105000 |
Specialist optics | Phase plate: VOLTA PHASE PLATE / Spherical aberration corrector: Krios with CS corrector / Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 14 eV |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 55.8 e/Å2 Details: 2678 images were collected in movie-mode with 60 frames per image on Gatan K3 in superresolution mode with 0.34 A pixel size / native pixel size 0.68A |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: RIGID BODY FIT |
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