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- EMDB-8918: Cryo-EM structure of FLNa ABD-E254K bound to phalloidin-stabilize... -
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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-8918 | |||||||||
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Title | Cryo-EM structure of FLNa ABD-E254K bound to phalloidin-stabilized F-actin, collected on a FEI Tecnai F20 microscope | |||||||||
![]() | Symmetrized FLNa-ABD-E254K bound to F-actin stabilized by phalloidin. Low-pass filtered to 7.4 Angstrom with a B-factor of -500. | |||||||||
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Biological species | ![]() ![]() ![]() | |||||||||
Method | helical reconstruction / cryo EM / Resolution: 7.4 Å | |||||||||
![]() | Iwamoto DV / Huehn AR / Simon B / Huet-Calderwoot C / Baldassarre M / Sindelar CV / Calderwood DA | |||||||||
![]() | ![]() Title: Structural basis of the filamin A actin-binding domain interaction with F-actin. Authors: Daniel V Iwamoto / Andrew Huehn / Bertrand Simon / Clotilde Huet-Calderwood / Massimiliano Baldassarre / Charles V Sindelar / David A Calderwood / ![]() ![]() Abstract: Actin-cross-linking proteins assemble actin filaments into higher-order structures essential for orchestrating cell shape, adhesion, and motility. Missense mutations in the tandem calponin homology ...Actin-cross-linking proteins assemble actin filaments into higher-order structures essential for orchestrating cell shape, adhesion, and motility. Missense mutations in the tandem calponin homology domains of their actin-binding domains (ABDs) underlie numerous genetic diseases, but a molecular understanding of these pathologies is hampered by the lack of high-resolution structures of any actin-cross-linking protein bound to F-actin. Here, taking advantage of a high-affinity, disease-associated mutant of the human filamin A (FLNa) ABD, we combine cryo-electron microscopy and functional studies to reveal at near-atomic resolution how the first calponin homology domain (CH1) and residues immediately N-terminal to it engage actin. We further show that reorientation of CH2 relative to CH1 is required to avoid clashes with actin and to expose F-actin-binding residues on CH1. Our data explain localization of disease-associated loss-of-function mutations to FLNaCH1 and gain-of-function mutations to the regulatory FLNaCH2. Sequence conservation argues that this provides a general model for ABD-F-actin binding. | |||||||||
History |
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Structure visualization
Movie |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 22.1 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 13.9 KB 13.9 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.9 KB | Display | ![]() |
Images | ![]() | 34.3 KB | ||
Masks | ![]() | 226.3 MB | ![]() | |
Others | ![]() ![]() | 32.6 MB 32.6 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 79 KB | Display | ![]() |
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Full document | ![]() | 78.1 KB | Display | |
Data in XML | ![]() | 495 B | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | Symmetrized FLNa-ABD-E254K bound to F-actin stabilized by phalloidin. Low-pass filtered to 7.4 Angstrom with a B-factor of -500. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.247 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
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Projections & Slices |
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Density Histograms |
-Half map: Symmetrized and unfiltered half map (1) of FLNa-ABD-E254K...
File | emd_8918_half_map_1.map | ||||||||||||
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Annotation | Symmetrized and unfiltered half map (1) of FLNa-ABD-E254K bound to F-actin stabilized by phalloidin | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Symmetrized and unfiltered half map (2) of FLNa-ABD-E254K...
File | emd_8918_half_map_2.map | ||||||||||||
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Annotation | Symmetrized and unfiltered half map (2) of FLNa-ABD-E254K bound to F-actin stabilized by phalloidin | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Helical filament of Filamin A actin-binding domain E254K bound to...
Entire | Name: Helical filament of Filamin A actin-binding domain E254K bound to F-actin stabilized by phalloidin |
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Components |
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-Supramolecule #1: Helical filament of Filamin A actin-binding domain E254K bound to...
Supramolecule | Name: Helical filament of Filamin A actin-binding domain E254K bound to F-actin stabilized by phalloidin type: complex / ID: 1 / Parent: 0 |
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-Supramolecule #2: Filamin A ABD-E254K
Supramolecule | Name: Filamin A ABD-E254K / type: complex / ID: 2 / Parent: 1 |
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Source (natural) | Organism: ![]() |
Recombinant expression | Organism: ![]() ![]() |
-Supramolecule #3: F-actin
Supramolecule | Name: F-actin / type: complex / ID: 3 / Parent: 1 |
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Source (natural) | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | helical reconstruction |
Aggregation state | helical array |
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Sample preparation
Buffer | pH: 7 |
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Vitrification | Cryogen name: ETHANE / Instrument: HOMEMADE PLUNGER |
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Electron microscopy
Microscope | FEI TECNAI F20 |
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Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 50.0 e/Å2 |
Electron beam | Acceleration voltage: 200 kV / Electron source: ![]() |
Electron optics | Illumination mode: SPOT SCAN / Imaging mode: BRIGHT FIELD |
Experimental equipment | ![]() Model: Tecnai F20 / Image courtesy: FEI Company |