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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-7832 | |||||||||
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Title | FLNaABD-Q170P bound to phalloidin-stabilized F-actin | |||||||||
![]() | Symmetrized map of FLNaABD-Q170P bound to phalloidin-stabilized F-actin. This map has been low-pass filtered to 6.6 A and sharpened with a B-factor of -250. | |||||||||
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Biological species | ![]() ![]() ![]() | |||||||||
Method | helical reconstruction / cryo EM / Resolution: 6.6 Å | |||||||||
![]() | Iwamoto DV / Huehn AR / Simon B / Huet-Calderwood 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 | ![]() | 20.8 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 14.6 KB 14.6 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 13.9 KB | Display | ![]() |
Images | ![]() | 34.1 KB | ||
Masks | ![]() | 226.3 MB | ![]() | |
Others | ![]() ![]() | 32.6 MB 32.6 MB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
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Links
EMDB pages | ![]() ![]() |
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Map
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Annotation | Symmetrized map of FLNaABD-Q170P bound to phalloidin-stabilized F-actin. This map has been low-pass filtered to 6.6 A and sharpened with a B-factor of -250. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: Independently refined half map(1) that was symmetrized with...
File | emd_7832_half_map_1.map | ||||||||||||
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Annotation | Independently refined half map(1) that was symmetrized with parameters derived from the full map | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Independently refined half map(2) that was symmetrized with...
File | emd_7832_half_map_2.map | ||||||||||||
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Annotation | Independently refined half map(2) that was symmetrized with parameters derived from the full map | ||||||||||||
Projections & Slices |
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Density Histograms |
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Sample components
-Entire : Helical complex of FLNaABD-Q170P bound to phalloidin-stabilized F...
Entire | Name: Helical complex of FLNaABD-Q170P bound to phalloidin-stabilized F-actin |
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Components |
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-Supramolecule #1: Helical complex of FLNaABD-Q170P bound to phalloidin-stabilized F...
Supramolecule | Name: Helical complex of FLNaABD-Q170P bound to phalloidin-stabilized F-actin type: complex / ID: 1 / Parent: 0 / Macromolecule list: all |
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Source (natural) | Organism: ![]() |
-Macromolecule #1: Filamin A Actin Binding Domain
Macromolecule | Name: Filamin A Actin Binding Domain / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() |
Sequence | String: PATEKDLAED APWKKIQQNT FTRWCNEHLK CVSKRIANLQ TDLSDGLRLI ALLEVLSQKK MHRKHNQRPT FRQMQLENVS VALEFLDRES IKLVSIDSKA IVDGNLKLIL GLIWTLILHY S |
-Macromolecule #2: Actin
Macromolecule | Name: Actin / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO |
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Source (natural) | Organism: ![]() ![]() |
Sequence | String: TTALVCDNGS GLVKAGFAGD DAPRAVFPSI VGRPRHQGVM VGMGQKDSYV GDEAQSKRGI LTLKYPIEHG IITNWDDMEK IWHHTFYNE LRVAPEEHPT LLTEAPLNPK ANREKMTQIM FETFNVPAMY VAIQAVLSLY ASGRTTGIVL DSGDGVTHNV P IYEGYALP ...String: TTALVCDNGS GLVKAGFAGD DAPRAVFPSI VGRPRHQGVM VGMGQKDSYV GDEAQSKRGI LTLKYPIEHG IITNWDDMEK IWHHTFYNE LRVAPEEHPT LLTEAPLNPK ANREKMTQIM FETFNVPAMY VAIQAVLSLY ASGRTTGIVL DSGDGVTHNV P IYEGYALP HAIMRLDLAG RDLTDYLMKI LTERGYSFVT TAEREIVRDI KEKLCYVALD FENEMATAAS SSSLEKSYEL PD GQVITIG NERFRCPETL FQPSFIGMES AGIHETTYNS IMKCDIDIRK DLYANNVMSG GTTMYPGIAD RMQKEITALA PST MKIKII APPERKYSVW IGGSILASLS TFQQMWITKQ EYDEAGPSIV HRKC |
-Experimental details
-Structure determination
Method | cryo EM |
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![]() | helical reconstruction |
Aggregation state | filament |
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Sample preparation
Buffer | pH: 8 |
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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: 60.0 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 |