+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-13991 | ||||||||||||||||||
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Title | In situ structure of actomyosin complex in skeletal sarcomere | ||||||||||||||||||
Map data | |||||||||||||||||||
Sample |
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Function / homology | Function and homology information skeletal muscle fiber adaptation / Striated Muscle Contraction / cellular response to organonitrogen compound / myosin filament / : / myosin II complex / response to steroid hormone / microfilament motor activity / myofibril / mesenchyme migration ...skeletal muscle fiber adaptation / Striated Muscle Contraction / cellular response to organonitrogen compound / myosin filament / : / myosin II complex / response to steroid hormone / microfilament motor activity / myofibril / mesenchyme migration / skeletal muscle thin filament assembly / striated muscle thin filament / skeletal muscle fiber development / response to mechanical stimulus / stress fiber / sarcomere / response to activity / muscle contraction / filopodium / actin filament / Hydrolases; Acting on acid anhydrides; Acting on acid anhydrides to facilitate cellular and subcellular movement / actin filament binding / actin cytoskeleton / double-stranded RNA binding / lamellipodium / cell body / calmodulin binding / hydrolase activity / positive regulation of gene expression / protein-containing complex / ATP binding / cytoplasm Similarity search - Function | ||||||||||||||||||
Biological species | Mus musculus (house mouse) / house mouse (house mouse) | ||||||||||||||||||
Method | subtomogram averaging / cryo EM / Resolution: 6.6 Å | ||||||||||||||||||
Authors | Wang Z / Grange M / Pospich S / Wagner T / Kho AL / Gautel M / Raunser S | ||||||||||||||||||
Funding support | European Union, 5 items
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Citation | Journal: Science / Year: 2022 Title: Structures from intact myofibrils reveal mechanism of thin filament regulation through nebulin. Authors: Zhexin Wang / Michael Grange / Sabrina Pospich / Thorsten Wagner / Ay Lin Kho / Mathias Gautel / Stefan Raunser / Abstract: In skeletal muscle, nebulin stabilizes and regulates the length of thin filaments, but the underlying mechanism remains nebulous. In this work, we used cryo-electron tomography and subtomogram ...In skeletal muscle, nebulin stabilizes and regulates the length of thin filaments, but the underlying mechanism remains nebulous. In this work, we used cryo-electron tomography and subtomogram averaging to reveal structures of native nebulin bound to thin filaments within intact sarcomeres. This in situ reconstruction provided high-resolution details of the interaction between nebulin and actin, demonstrating the stabilizing role of nebulin. Myosin bound to the thin filaments exhibited different conformations of the neck domain, highlighting its inherent structural variability in muscle. Unexpectedly, nebulin did not interact with myosin or tropomyosin, but it did interact with a troponin T linker through two potential binding motifs on nebulin, explaining its regulatory role. Our structures support the role of nebulin as a thin filament "molecular ruler" and provide a molecular basis for studying nemaline myopathies. | ||||||||||||||||||
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_13991.map.gz | 3.8 MB | EMDB map data format | |
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Header (meta data) | emd-13991-v30.xml emd-13991.xml | 25.6 KB 25.6 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_13991_fsc.xml | 9.2 KB | Display | FSC data file |
Images | emd_13991.png | 93.1 KB | ||
Masks | emd_13991_msk_1.map | 30.5 MB | Mask map | |
Others | emd_13991_half_map_1.map.gz emd_13991_half_map_2.map.gz | 28.3 MB 28.3 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-13991 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-13991 | HTTPS FTP |
-Related structure data
Related structure data | 7qinMC 7qimC 7qioC C: citing same article (ref.) M: atomic model generated by this map |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_13991.map.gz / Format: CCP4 / Size: 30.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.73 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Mask #1
File | emd_13991_msk_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #2
File | emd_13991_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_13991_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Mouse psoas muscle myofibrils
+Supramolecule #1: Mouse psoas muscle myofibrils
+Macromolecule #1: Actin, alpha skeletal muscle
+Macromolecule #2: nebulin (mouse)
+Macromolecule #3: nebulin (mouse)
+Macromolecule #4: tropomyosin, alpha-1 (mouse)
+Macromolecule #5: tropomyosin, alpha-1 (mouse)
+Macromolecule #6: tropomyosin, alpha-1 (mouse)
+Macromolecule #7: Myosin-4
+Macromolecule #8: ADENOSINE-5'-DIPHOSPHATE
+Macromolecule #9: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | subtomogram averaging |
Aggregation state | tissue |
-Sample preparation
Buffer | pH: 7 |
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Vitrification | Cryogen name: ETHANE / Instrument: FEI VITROBOT MARK IV |
-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 / Calibrated defocus max: 5.0 µm / Calibrated defocus min: 2.4 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal magnification: 81000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Average electron dose: 3.4 e/Å2 |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |