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Yorodumi- EMDB-22712: Subtomogram average of flagellar axoneme doublet from the wild ty... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-22712 | |||||||||||||||
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Title | Subtomogram average of flagellar axoneme doublet from the wild type Tetrahymena thermophila | |||||||||||||||
Map data | Subtomogram average of flagellar axoneme doublet from the wild type Tetrahymena thermophila | |||||||||||||||
Sample |
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Biological species | Tetrahymena thermophila (eukaryote) | |||||||||||||||
Method | subtomogram averaging / cryo EM / Resolution: 17.3 Å | |||||||||||||||
Authors | Fabritius AS / Bayless BA / Li S / Stoddard D / Heydeck W / Ebmeier CC / Anderson L / Gunnels T / Nachiappan C / Whitall J ...Fabritius AS / Bayless BA / Li S / Stoddard D / Heydeck W / Ebmeier CC / Anderson L / Gunnels T / Nachiappan C / Whitall J / Old W / Agard DA / Nicastro D / Winey M | |||||||||||||||
Funding support | United States, 4 items
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Citation | Journal: Mol Biol Cell / Year: 2021 Title: Proteomic analysis of microtubule inner proteins (MIPs) in Rib72 null cells reveals functional MIPs. Authors: Amy S Fabritius / Brian A Bayless / Sam Li / Daniel Stoddard / Westley Heydeck / Christopher C Ebmeier / Lauren Anderson / Tess Gunnels / Chidambaram Nachiappan / Justen B Whittall / William ...Authors: Amy S Fabritius / Brian A Bayless / Sam Li / Daniel Stoddard / Westley Heydeck / Christopher C Ebmeier / Lauren Anderson / Tess Gunnels / Chidambaram Nachiappan / Justen B Whittall / William Old / David A Agard / Daniela Nicastro / Mark Winey / Abstract: The core structure of motile cilia and flagella, the axoneme, is built from a stable population of doublet microtubules. This unique stability is brought about, at least in part, by a network of ...The core structure of motile cilia and flagella, the axoneme, is built from a stable population of doublet microtubules. This unique stability is brought about, at least in part, by a network of microtubule inner proteins (MIPs) that are bound to the luminal side of the microtubule walls. Rib72A and Rib72B were identified as MIPs in the motile cilia of the protist . Loss of these proteins leads to ciliary defects and loss of additional MIPs. We performed mass spectrometry coupled with proteomic analysis and bioinformatics to identify the MIPs lost in knockout axonemes. We identified a number of candidate MIPs and pursued one, Fap115, for functional characterization. We find that loss of Fap115 results in disrupted cell swimming and aberrant ciliary beating. Cryo-electron tomography reveals that Fap115 localizes to MIP6a in the A-tubule of the doublet microtubules. Overall, our results highlight the complex relationship between MIPs, ciliary structure, and ciliary function. | |||||||||||||||
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_22712.map.gz | 9.9 MB | EMDB map data format | |
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Header (meta data) | emd-22712-v30.xml emd-22712.xml | 9.9 KB 9.9 KB | Display Display | EMDB header |
Images | emd_22712.png | 90.2 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-22712 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-22712 | HTTPS FTP |
-Validation report
Summary document | emd_22712_validation.pdf.gz | 320.5 KB | Display | EMDB validaton report |
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Full document | emd_22712_full_validation.pdf.gz | 320.1 KB | Display | |
Data in XML | emd_22712_validation.xml.gz | 5.6 KB | Display | |
Data in CIF | emd_22712_validation.cif.gz | 6.4 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22712 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-22712 | HTTPS FTP |
-Related structure data
Related structure data | C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_22712.map.gz / Format: CCP4 / Size: 10.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Subtomogram average of flagellar axoneme doublet from the wild type Tetrahymena thermophila | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 5.3 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
-Entire : flagellum
Entire | Name: flagellum |
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Components |
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-Supramolecule #1: flagellum
Supramolecule | Name: flagellum / type: organelle_or_cellular_component / ID: 1 / Parent: 0 Details: flagellar axoneme isolated from Tetrahymena thermophila |
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Source (natural) | Organism: Tetrahymena thermophila (eukaryote) |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | cell |
-Sample preparation
Buffer | pH: 7 |
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Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Average electron dose: 80.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Applied symmetry - Point group: C1 (asymmetric) / Algorithm: BACK PROJECTION / Resolution.type: BY AUTHOR / Resolution: 17.3 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: Xmipp (ver. 2.4) / Number subtomograms used: 5235 |
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Extraction | Number tomograms: 5235 / Number images used: 5235 |
Final angle assignment | Type: MAXIMUM LIKELIHOOD |