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Yorodumi- EMDB-51015: Structure of the Position 8 to 9 CMG-decorated gamma-Tubulin Ring... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-51015 | |||||||||
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Title | Structure of the Position 8 to 9 CMG-decorated gamma-Tubulin Ring Complex from Pig Brain | |||||||||
Map data | ||||||||||
Sample |
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Keywords | Tubulin Complex / STRUCTURAL PROTEIN | |||||||||
Biological species | Sus scrofa (pig) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.6 Å | |||||||||
Authors | Munoz-Hernandez H / Wieczorek M | |||||||||
Funding support | Switzerland, 2 items
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Citation | Journal: Dev Cell / Year: 2024 Title: Partial closure of the γ-tubulin ring complex by CDK5RAP2 activates microtubule nucleation. Authors: Yixin Xu / Hugo Muñoz-Hernández / Rościsław Krutyhołowa / Florina Marxer / Ferdane Cetin / Michal Wieczorek / Abstract: Microtubule nucleation is templated by the γ-tubulin ring complex (γ-TuRC), but its structure deviates from the geometry of α-/β-tubulin in the microtubule, explaining the complex's poor ...Microtubule nucleation is templated by the γ-tubulin ring complex (γ-TuRC), but its structure deviates from the geometry of α-/β-tubulin in the microtubule, explaining the complex's poor nucleating activity. Several proteins may activate the γ-TuRC, but the mechanisms underlying activation are not known. Here, we determined the structure of the porcine γ-TuRC purified using CDK5RAP2's centrosomin motif 1 (CM1). We identified an unexpected conformation of the γ-TuRC bound to multiple protein modules containing MZT2, GCP2, and CDK5RAP2, resulting in a long-range constriction of the γ-tubulin ring that brings it in closer agreement with the 13-protofilament microtubule. Additional CDK5RAP2 promoted γ-TuRC decoration and stimulated the microtubule-nucleating activities of the porcine γ-TuRC and a reconstituted, CM1-free human complex in single-molecule assays. Our results provide a structural mechanism for the control of microtubule nucleation by CM1 proteins and identify conformational transitions in the γ-TuRC that prime it for microtubule nucleation. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_51015.map.gz | 148.2 MB | EMDB map data format | |
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Header (meta data) | emd-51015-v30.xml emd-51015.xml | 15.3 KB 15.3 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_51015_fsc.xml | 13.6 KB | Display | FSC data file |
Images | emd_51015.png | 59.2 KB | ||
Filedesc metadata | emd-51015.cif.gz | 3.8 KB | ||
Others | emd_51015_half_map_1.map.gz emd_51015_half_map_2.map.gz | 136.7 MB 136.7 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-51015 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-51015 | HTTPS FTP |
-Validation report
Summary document | emd_51015_validation.pdf.gz | 762.4 KB | Display | EMDB validaton report |
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Full document | emd_51015_full_validation.pdf.gz | 761.9 KB | Display | |
Data in XML | emd_51015_validation.xml.gz | 20.8 KB | Display | |
Data in CIF | emd_51015_validation.cif.gz | 27.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-51015 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-51015 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_51015.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.4133 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #1
File | emd_51015_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_51015_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
-Entire : Gamma-Tubulin Ring Complex from native pig brain
Entire | Name: Gamma-Tubulin Ring Complex from native pig brain |
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Components |
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-Supramolecule #1: Gamma-Tubulin Ring Complex from native pig brain
Supramolecule | Name: Gamma-Tubulin Ring Complex from native pig brain / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#24 |
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Source (natural) | Organism: Sus scrofa (pig) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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Grid | Model: Quantifoil R2/1 / Support film - Material: CARBON / Support film - topology: HOLEY |
Vitrification | Cryogen name: ETHANE-PROPANE |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 55.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.9 µm / Nominal defocus min: 0.9 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: BACKBONE TRACE |
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