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Yorodumi- EMDB-50845: Real space helical reconstruction of cofilin actin in the microtu... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-50845 | |||||||||||||||||||||||||||||||||||||||
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Title | Real space helical reconstruction of cofilin actin in the microtubule lumen of HAP1 cells | |||||||||||||||||||||||||||||||||||||||
Map data | Real space helical reconstruction of cofilin actin in the microtubule lumen of HAP1 cells | |||||||||||||||||||||||||||||||||||||||
Sample |
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Keywords | cofilin / actin / filament / CYTOSOLIC PROTEIN | |||||||||||||||||||||||||||||||||||||||
Biological species | Homo sapiens (human) | |||||||||||||||||||||||||||||||||||||||
Method | helical reconstruction / cryo EM / Resolution: 20.0 Å | |||||||||||||||||||||||||||||||||||||||
Authors | Tsuji C / Bradshaw M / Paul DM / Dodding MP | |||||||||||||||||||||||||||||||||||||||
Funding support | United Kingdom, 12 items
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Citation | Journal: Nat Commun / Year: 2024 Title: CryoET reveals actin filaments within platelet microtubules. Authors: Chisato Tsuji / Marston Bradshaw / Megan F Allen / Molly L Jackson / Judith Mantell / Ufuk Borucu / Alastair W Poole / Paul Verkade / Ingeborg Hers / Danielle M Paul / Mark P Dodding / Abstract: Crosstalk between the actin and microtubule cytoskeletons is important for many cellular processes. Recent studies have shown that microtubules and F-actin can assemble to form a composite structure ...Crosstalk between the actin and microtubule cytoskeletons is important for many cellular processes. Recent studies have shown that microtubules and F-actin can assemble to form a composite structure where F-actin occupies the microtubule lumen. Whether these cytoskeletal hybrids exist in physiological settings and how they are formed is unclear. Here, we show that the short-crossover Class I actin filament previously identified inside microtubules in human HAP1 cells is cofilin-bound F-actin. Lumenal F-actin can be reconstituted in vitro, but cofilin is not essential. Moreover, actin filaments with both cofilin-bound and canonical morphologies reside within human platelet microtubules under physiological conditions. We propose that stress placed upon the microtubule network during motor-driven microtubule looping and sliding may facilitate the incorporation of actin into microtubules. | |||||||||||||||||||||||||||||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_50845.map.gz | 7.6 MB | EMDB map data format | |
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Header (meta data) | emd-50845-v30.xml emd-50845.xml | 11.3 KB 11.3 KB | Display Display | EMDB header |
Images | emd_50845.png | 26.7 KB | ||
Filedesc metadata | emd-50845.cif.gz | 4.1 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-50845 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-50845 | HTTPS FTP |
-Validation report
Summary document | emd_50845_validation.pdf.gz | 325.9 KB | Display | EMDB validaton report |
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Full document | emd_50845_full_validation.pdf.gz | 325.4 KB | Display | |
Data in XML | emd_50845_validation.xml.gz | 5.1 KB | Display | |
Data in CIF | emd_50845_validation.cif.gz | 5.7 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-50845 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-50845 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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-Map
File | Download / File: emd_50845.map.gz / Format: CCP4 / Size: 8 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Annotation | Real space helical reconstruction of cofilin actin in the microtubule lumen of HAP1 cells | ||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 3.75 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Sample components
-Entire : Cofilin bound actin structure found in the lumen of microtubules ...
Entire | Name: Cofilin bound actin structure found in the lumen of microtubules in HAP1 cells |
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Components |
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-Supramolecule #1: Cofilin bound actin structure found in the lumen of microtubules ...
Supramolecule | Name: Cofilin bound actin structure found in the lumen of microtubules in HAP1 cells type: complex / ID: 1 / Parent: 0 |
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Source (natural) | Organism: Homo sapiens (human) / Strain: HAP1 cells |
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | helical reconstruction |
Aggregation state | filament |
-Sample preparation
Buffer | pH: 6.8 |
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Vitrification | Cryogen name: ETHANE / Instrument: LEICA EM GP |
-Electron microscopy
Microscope | TFS KRIOS |
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Image recording | Film or detector model: TFS FALCON 4i (4k x 4k) / Average electron dose: 2.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm / Nominal defocus max: 7.0 µm / Nominal defocus min: 3.0 µm / Nominal magnification: 63000 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Final reconstruction | Applied symmetry - Helical parameters - Δz: 27.5 Å Applied symmetry - Helical parameters - Δ&Phi: 162 ° Applied symmetry - Helical parameters - Axial symmetry: C1 (asymmetric) Resolution.type: BY AUTHOR / Resolution: 20.0 Å / Resolution method: OTHER Details: A sub-volume containing the filament was extracted from a tomogram, which was then 2D projected. The projected image was used for real space helical reconstruction to create the map. ...Details: A sub-volume containing the filament was extracted from a tomogram, which was then 2D projected. The projected image was used for real space helical reconstruction to create the map. Resolution was estimated compared to molmap generation of PDB:3J0S at 20 angstroms in Chimera and correlation score of 0.96 was obtained. Number images used: 1 |
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Startup model | Type of model: NONE |
Final angle assignment | Type: NOT APPLICABLE |