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Yorodumi- EMDB-19496: Structure of the formin Cdc12 bound to the barbed end of phalloid... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-19496 | ||||||||||||
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Title | Structure of the formin Cdc12 bound to the barbed end of phalloidin-stabilized F-actin. | ||||||||||||
Map data | Sharpened, local-resolution filtered cryo-EM density map of the formin Cdc12 bound to the barbed end of phalloidin-stabilized F-actin. | ||||||||||||
Sample |
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Keywords | actin / formin / Cdc12 / actin assembly. / STRUCTURAL PROTEIN | ||||||||||||
Function / homology | Function and homology information F-bar domain binding / protein localization to mitotic actomyosin contractile ring / medial cortical node / mitotic actomyosin contractile ring, proximal layer / mitotic actomyosin contractile ring / medial cortex / mitotic actomyosin contractile ring assembly / positive regulation of norepinephrine uptake / cellular response to cytochalasin B / bBAF complex ...F-bar domain binding / protein localization to mitotic actomyosin contractile ring / medial cortical node / mitotic actomyosin contractile ring, proximal layer / mitotic actomyosin contractile ring / medial cortex / mitotic actomyosin contractile ring assembly / positive regulation of norepinephrine uptake / cellular response to cytochalasin B / bBAF complex / npBAF complex / postsynaptic actin cytoskeleton organization / regulation of transepithelial transport / brahma complex / nBAF complex / structural constituent of postsynaptic actin cytoskeleton / morphogenesis of a polarized epithelium / Formation of annular gap junctions / GBAF complex / Gap junction degradation / postsynaptic actin cytoskeleton / protein localization to adherens junction / regulation of G0 to G1 transition / dense body / Cell-extracellular matrix interactions / Tat protein binding / Folding of actin by CCT/TriC / regulation of double-strand break repair / regulation of nucleotide-excision repair / RSC-type complex / apical protein localization / mating projection tip / Prefoldin mediated transfer of substrate to CCT/TriC / barbed-end actin filament capping / adherens junction assembly / RHOF GTPase cycle / Adherens junctions interactions / tight junction / Sensory processing of sound by outer hair cells of the cochlea / SWI/SNF complex / Interaction between L1 and Ankyrins / Sensory processing of sound by inner hair cells of the cochlea / regulation of mitotic metaphase/anaphase transition / regulation of norepinephrine uptake / positive regulation of double-strand break repair / positive regulation of T cell differentiation / NuA4 histone acetyltransferase complex / regulation of synaptic vesicle endocytosis / apical junction complex / maintenance of blood-brain barrier / establishment or maintenance of cell polarity / cell division site / cortical cytoskeleton / positive regulation of double-strand break repair via homologous recombination / positive regulation of stem cell population maintenance / nitric-oxide synthase binding / Recycling pathway of L1 / regulation of cyclin-dependent protein serine/threonine kinase activity / regulation of G1/S transition of mitotic cell cycle / actin filament bundle assembly / negative regulation of cell differentiation / brush border / kinesin binding / calyx of Held / EPH-ephrin mediated repulsion of cells / RHO GTPases Activate WASPs and WAVEs / RHO GTPases activate IQGAPs / positive regulation of myoblast differentiation / regulation of protein localization to plasma membrane / EPHB-mediated forward signaling / substantia nigra development / actin filament polymerization / axonogenesis / negative regulation of protein binding / actin filament / cell motility / RHO GTPases Activate Formins / Translocation of SLC2A4 (GLUT4) to the plasma membrane / regulation of transmembrane transporter activity / positive regulation of cell differentiation / FCGR3A-mediated phagocytosis / adherens junction / Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement / DNA Damage Recognition in GG-NER / tau protein binding / Signaling by high-kinase activity BRAF mutants / Schaffer collateral - CA1 synapse / MAP2K and MAPK activation / B-WICH complex positively regulates rRNA expression / structural constituent of cytoskeleton / cytoplasmic ribonucleoprotein granule / kinetochore / Regulation of actin dynamics for phagocytic cup formation / platelet aggregation / small GTPase binding / nuclear matrix / VEGFA-VEGFR2 Pathway / UCH proteinases / Signaling by RAF1 mutants / Signaling by moderate kinase activity BRAF mutants Similarity search - Function | ||||||||||||
Biological species | Homo sapiens (human) / Schizosaccharomyces pombe (fission yeast) / Amanita phalloides (death cap) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.56 Å | ||||||||||||
Authors | Oosterheert W / Boiero Sanders M / Funk J / Prumbaum D / Raunser S / Bieling P | ||||||||||||
Funding support | Germany, European Union, 3 items
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Citation | Journal: To Be Published Title: Mechanism of formin-mediated processive elongation of actin filaments revealed by cryo-EM Authors: Oosterheert W / Boiero Sanders M / Funk J / Prumbaum D / Raunser S / Bieling P | ||||||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_19496.map.gz | 77.9 MB | EMDB map data format | |
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Header (meta data) | emd-19496-v30.xml emd-19496.xml | 38.8 KB 38.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_19496_fsc.xml | 11.4 KB | Display | FSC data file |
Images | emd_19496.png | 90.3 KB | ||
Masks | emd_19496_msk_1.map | 125 MB | Mask map | |
Filedesc metadata | emd-19496.cif.gz | 8.5 KB | ||
Others | emd_19496_additional_1.map.gz emd_19496_additional_2.map.gz emd_19496_additional_3.map.gz emd_19496_additional_4.map.gz emd_19496_additional_5.map.gz emd_19496_additional_6.map.gz emd_19496_half_map_1.map.gz emd_19496_half_map_2.map.gz | 98.3 MB 97.4 MB 75.9 MB 98.4 MB 98.4 MB 77.2 MB 98.5 MB 98.4 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-19496 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-19496 | HTTPS FTP |
-Related structure data
Related structure data | 8rttMC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_19496.map.gz / Format: CCP4 / Size: 125 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
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Annotation | Sharpened, local-resolution filtered cryo-EM density map of the formin Cdc12 bound to the barbed end of phalloidin-stabilized F-actin. | ||||||||||||||||||||
Voxel size | X=Y=Z: 0.88 Å | ||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Mask #1
File | emd_19496_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Additional map: 3D-refined, unsharpened cryo-EM density map of the formin...
File | emd_19496_additional_1.map | ||||||||||||
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Annotation | 3D-refined, unsharpened cryo-EM density map of the formin Cdc12 bound to the barbed end of phalloidin-stabilized F-actin. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Unsharpened cryo-EM density map of actin-Cdc12. This reconstruction...
File | emd_19496_additional_2.map | ||||||||||||
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Annotation | Unsharpened cryo-EM density map of actin-Cdc12. This reconstruction was computed with more particles, but displays weaker density for the FH2L domain of Cdc12 due to flexibility. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Composite map of two reconstructions of the formin...
File | emd_19496_additional_3.map | ||||||||||||
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Annotation | Composite map of two reconstructions of the formin Cdc12 bound to the barbed end of phalloidin-stabilized F-actin. This map was created using phenix and was used for visualization purposes. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Unfiltered half map 1 of actin-Cdc12. This reconstruction...
File | emd_19496_additional_4.map | ||||||||||||
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Annotation | Unfiltered half map 1 of actin-Cdc12. This reconstruction was computed with more particles, but displays weaker density for the FH2L domain of Cdc12 due to flexibility. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Unfiltered half map 2 of actin-Cdc12. This reconstruction...
File | emd_19496_additional_5.map | ||||||||||||
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Annotation | Unfiltered half map 2 of actin-Cdc12. This reconstruction was computed with more particles, but displays weaker density for the FH2L domain of Cdc12 due to flexibility. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Additional map: Sharpened density map of actin-Cdc12. This reconstruction was...
File | emd_19496_additional_6.map | ||||||||||||
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Annotation | Sharpened density map of actin-Cdc12. This reconstruction was computed with more particles, but displays weaker density for the FH2L domain of Cdc12 due to flexibility. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Unfiltered half map 1 of the formin Cdc12...
File | emd_19496_half_map_1.map | ||||||||||||
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Annotation | Unfiltered half map 1 of the formin Cdc12 bound to the barbed end of phalloidin-stabilized F-actin. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Half map: Unfiltered half map 2 of the formin Cdc12...
File | emd_19496_half_map_2.map | ||||||||||||
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Annotation | Unfiltered half map 2 of the formin Cdc12 bound to the barbed end of phalloidin-stabilized F-actin. | ||||||||||||
Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Complex of the dimeric FH2 domain of S. Pombe Cdc12 bound to the ...
+Supramolecule #1: Complex of the dimeric FH2 domain of S. Pombe Cdc12 bound to the ...
+Supramolecule #2: Actin filament
+Supramolecule #3: Dimeric FH2 domain of S. Pombe Cdc12
+Supramolecule #4: Phalloidin.
+Macromolecule #1: Actin, cytoplasmic 1, N-terminally processed
+Macromolecule #2: Cell division control protein 12
+Macromolecule #3: Phalloidin
+Macromolecule #4: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #5: MAGNESIUM ION
+Macromolecule #6: PHOSPHATE ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.1 Component:
Details: 1xKMEH (10 mM HEPES pH 7.1, 100 mM KCl, 2 mM MgCl2, 1 mM EGTA, 0.5 mM TCEP) | ||||||||||||||||||
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Grid | Model: Quantifoil R2/1 / Material: GOLD / Mesh: 200 / Support film - Material: CARBON / Support film - topology: HOLEY / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 90 sec. | ||||||||||||||||||
Vitrification | Cryogen name: ETHANE-PROPANE / Chamber humidity: 100 % / Chamber temperature: 286 K / Instrument: FEI VITROBOT MARK IV / Details: 3 seconds, force 0.. |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | C2 aperture diameter: 50.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 0.01 mm / Nominal defocus max: 2.7 µm / Nominal defocus min: 1.2 µm / Nominal magnification: 81000 |
Specialist optics | Spherical aberration corrector: Titan Krios G2 microscope (Thermo Fisher Scientific) with an in-column Cs-corrector. Energy filter - Name: GIF Bioquantum / Energy filter - Slit width: 15 eV / Details: Gatan energy filter. |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Details | 300 kV Titan Krios G2 microscope (Thermo Fisher Scientific) with an in-column Cs-corrector. |
Image recording | Film or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Number grids imaged: 1 / Number real images: 20393 / Average electron dose: 64.6 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
-Atomic model buiding 1
Initial model |
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Refinement | Space: REAL / Protocol: RIGID BODY FIT | ||||||||||||
Output model | PDB-8rtt: |