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Yorodumi- EMDB-20631: Composite cryo-EM density map of the 48-nm repeat doublet microtu... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-20631 | |||||||||
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Title | Composite cryo-EM density map of the 48-nm repeat doublet microtubule from Chlamydomonas reinhardtii | |||||||||
Map data | Composite cryo-EM density map of the 48-nm repeat doublet microtubule from Chlamydomonas reinhardtii | |||||||||
Sample |
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Keywords | motile cilia / microtubule doublet / repetitive structure / PROTEIN FIBRIL | |||||||||
Function / homology | Function and homology information axonemal central pair / axonemal doublet microtubule / positive regulation of cilium-dependent cell motility / outer dynein arm / outer dynein arm assembly / cilium-dependent cell motility / membrane-bounded organelle / cilium movement involved in cell motility / regulation of cilium beat frequency involved in ciliary motility / establishment of protein localization to organelle ...axonemal central pair / axonemal doublet microtubule / positive regulation of cilium-dependent cell motility / outer dynein arm / outer dynein arm assembly / cilium-dependent cell motility / membrane-bounded organelle / cilium movement involved in cell motility / regulation of cilium beat frequency involved in ciliary motility / establishment of protein localization to organelle / cilium movement / axoneme assembly / axonemal microtubule / cilium organization / negative regulation of microtubule depolymerization / nucleoside-diphosphate kinase / UTP biosynthetic process / CTP biosynthetic process / motile cilium / GTP biosynthetic process / microtubule associated complex / nucleoside diphosphate kinase activity / intracellular membraneless organelle / axoneme / microtubule-based process / mitotic cytokinesis / cilium assembly / alpha-tubulin binding / Hsp70 protein binding / mitotic spindle organization / ciliary basal body / meiotic cell cycle / ATP-dependent protein folding chaperone / Hsp90 protein binding / cilium / structural constituent of cytoskeleton / mitotic spindle / Hydrolases; Acting on acid anhydrides; Acting on GTP to facilitate cellular and subcellular movement / microtubule / calmodulin binding / hydrolase activity / GTPase activity / calcium ion binding / GTP binding / ATP binding / nucleus / metal ion binding / cytoplasm Similarity search - Function | |||||||||
Biological species | Chlamydomonas reinhardtii (plant) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.4 Å | |||||||||
Authors | Ma M / Stoyanova M / Rademacher G / Dutcher SK / Brown A / Zhang R | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Cell / Year: 2019 Title: Structure of the Decorated Ciliary Doublet Microtubule. Authors: Meisheng Ma / Mihaela Stoyanova / Griffin Rademacher / Susan K Dutcher / Alan Brown / Rui Zhang / Abstract: The axoneme of motile cilia is the largest macromolecular machine of eukaryotic cells. In humans, impaired axoneme function causes a range of ciliopathies. Axoneme assembly, structure, and motility ...The axoneme of motile cilia is the largest macromolecular machine of eukaryotic cells. In humans, impaired axoneme function causes a range of ciliopathies. Axoneme assembly, structure, and motility require a radially arranged set of doublet microtubules, each decorated in repeating patterns with non-tubulin components. We use single-particle cryo-electron microscopy to visualize and build an atomic model of the repeating structure of a native axonemal doublet microtubule, which reveals the identities, positions, repeat lengths, and interactions of 38 associated proteins, including 33 microtubule inner proteins (MIPs). The structure demonstrates how these proteins establish the unique architecture of doublet microtubules, maintain coherent periodicities along the axoneme, and stabilize the microtubules against the repeated mechanical stress induced by ciliary motility. Our work elucidates the architectural principles that underpin the assembly of this large, repetitive eukaryotic structure and provides a molecular basis for understanding the etiology of human ciliopathies. | |||||||||
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_20631.map.gz | 204.4 MB | EMDB map data format | |
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Header (meta data) | emd-20631-v30.xml emd-20631.xml | 63.7 KB 63.7 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_20631_fsc.xml | 18 KB | Display | FSC data file |
Images | emd_20631.png | 149.7 KB | ||
Masks | emd_20631_msk_1.map | 512 MB | Mask map | |
Filedesc metadata | emd-20631.cif.gz | 15.7 KB | ||
Others | emd_20631_half_map_1.map.gz emd_20631_half_map_2.map.gz | 173.6 MB 173 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-20631 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-20631 | HTTPS FTP |
-Validation report
Summary document | emd_20631_validation.pdf.gz | 1 MB | Display | EMDB validaton report |
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Full document | emd_20631_full_validation.pdf.gz | 1 MB | Display | |
Data in XML | emd_20631_validation.xml.gz | 25.8 KB | Display | |
Data in CIF | emd_20631_validation.cif.gz | 34.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20631 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-20631 | HTTPS FTP |
-Related structure data
Related structure data | 6u42MC 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_20631.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Composite cryo-EM density map of the 48-nm repeat doublet microtubule from Chlamydomonas reinhardtii | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.403 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_20631_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: Composite cryo-EM density map of the 48-nm repeat...
File | emd_20631_half_map_1.map | ||||||||||||
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Annotation | Composite cryo-EM density map of the 48-nm repeat doublet microtubule from Chlamydomonas reinhardtii (half map 1) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Composite cryo-EM density map of the 48-nm repeat...
File | emd_20631_half_map_2.map | ||||||||||||
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Annotation | Composite cryo-EM density map of the 48-nm repeat doublet microtubule from Chlamydomonas reinhardtii (half map 2) | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Doublet microtubule from wild-type Chlamydomonas reinhardtii
+Supramolecule #1: Doublet microtubule from wild-type Chlamydomonas reinhardtii
+Macromolecule #1: Tubulin beta
+Macromolecule #2: Tubulin alpha
+Macromolecule #3: PACRG
+Macromolecule #4: FAP20
+Macromolecule #5: FAP53
+Macromolecule #6: FAP127
+Macromolecule #7: RIB43a
+Macromolecule #8: FAP112
+Macromolecule #9: FAP210
+Macromolecule #10: FAP45
+Macromolecule #11: RIB30
+Macromolecule #12: FAP363
+Macromolecule #13: FAP126
+Macromolecule #14: FAP52
+Macromolecule #15: FAP67
+Macromolecule #16: FAP85
+Macromolecule #17: FAP106
+Macromolecule #18: FAP115
+Macromolecule #19: FAP143
+Macromolecule #20: FAP161
+Macromolecule #21: FAP166
+Macromolecule #22: FAP252
+Macromolecule #23: FAP276
+Macromolecule #24: RIB72
+Macromolecule #25: FAP141
+Macromolecule #26: FAP222
+Macromolecule #27: FAP95
+Macromolecule #28: FAP182
+Macromolecule #29: FAP129
+Macromolecule #30: FAP21
+Macromolecule #31: FAP273
+Macromolecule #32: FAP107
+Macromolecule #33: RIB21
+Macromolecule #34: DC1
+Macromolecule #35: DC2
+Macromolecule #36: DC3
+Macromolecule #37: FAP68
+Macromolecule #38: FAP90
+Macromolecule #39: GUANOSINE-5'-DIPHOSPHATE
+Macromolecule #40: MAGNESIUM ION
+Macromolecule #41: GUANOSINE-5'-TRIPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | filament |
-Sample preparation
Buffer | pH: 7.4 / Component - Name: HMDEKP Details: 30 mM HEPES, 5 mM MgSO4, 1 mM DTT, 0.5 mM EGTA, 25 mM KCl, PH 7.4 |
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Vitrification | Cryogen name: ETHANE / Chamber humidity: 95 % / Chamber temperature: 277 K / Instrument: FEI VITROBOT MARK IV / Details: blot for 4 seconds before plunging. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Specialist optics | Spherical aberration corrector: Microscope is equipped with a Cs corrector Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 20 eV |
Image recording | Film or detector model: GATAN K2 QUANTUM (4k x 4k) / Detector mode: COUNTING / Digitization - Dimensions - Width: 3838 pixel / Digitization - Dimensions - Height: 3710 pixel / Digitization - Frames/image: 1-30 / Number grids imaged: 6 / Number real images: 8314 / Average exposure time: 9.0 sec. / Average electron dose: 38.9 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Calibrated defocus max: 3.1 µm / Calibrated defocus min: 1.0 µm / Calibrated magnification: 81000 / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 0.01 mm / Nominal defocus max: 2.75 µm / Nominal defocus min: 1.25 µm / Nominal magnification: 81000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: REAL / Protocol: AB INITIO MODEL / Overall B value: 50 |
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Output model | PDB-6u42: |