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TitleStructures from intact myofibrils reveal mechanism of thin filament regulation through nebulin.
Journal, issue, pagesScience, Vol. 375, Issue 6582, Page eabn1934, Year 2022
Publish dateFeb 18, 2022
AuthorsZhexin Wang / Michael Grange / Sabrina Pospich / Thorsten Wagner / Ay Lin Kho / Mathias Gautel / Stefan Raunser /
PubMed AbstractIn 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.
External linksScience / PubMed:35175800
MethodsEM (subtomogram averaging) / EM (tomography)
Resolution4.5 - 12.0 Å
Structure data

EMDB-13990, PDB-7qim:
In situ structure of nebulin bound to actin filament in skeletal sarcomere
Method: EM (subtomogram averaging) / Resolution: 4.5 Å

EMDB-13991, PDB-7qin:
In situ structure of actomyosin complex in skeletal sarcomere
Method: EM (subtomogram averaging) / Resolution: 6.6 Å

EMDB-13992:
In situ structure of myosin neck domain in skeletal sarcomere (centered on essential light chain)
Method: EM (subtomogram averaging) / Resolution: 8.9 Å

EMDB-13993: In situ structure of myosin neck domain in skeletal sarcomere (centered on regulatory light chain)
PDB-7qio: Homology model of myosin neck domain in skeletal sarcomere
Method: EM (subtomogram averaging) / Resolution: 9.0 Å

EMDB-13994:
In situ structure of nebulin bound to F-actin in skeletal sarcomere I-band
Method: EM (subtomogram averaging) / Resolution: 7.4 Å

EMDB-13995:
In situ structure of F-actin in thin filament from cardiac sarcomere
Method: EM (subtomogram averaging) / Resolution: 6.3 Å

EMDB-13996:
In situ structure of actomyosin in cardiac sarcomere
Method: EM (subtomogram averaging) / Resolution: 7.7 Å

EMDB-13997:
In situ structure of myosin double-head in cardiac sarcomere
Method: EM (subtomogram averaging) / Resolution: 12.0 Å

EMDB-13998:
Tomogram of skeletal sarcomere A-band after FIB-milling
Method: EM (tomography)

EMDB-13999:
Tomogram of skeletal sarcomere I-band after FIB-milling
Method: EM (tomography)

EMDB-14000:
Tomogram of cardiac sarcomere A-band and M-band after FIB-milling
Method: EM (tomography)

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM / Adenosine diphosphate

ChemComp-MG:
Unknown entry

Source
  • mus musculus (house mouse)
  • house mouse (house mouse)
KeywordsCONTRACTILE PROTEIN / Skeletal muscle / Sarcomere / Thin filament / Cross-bridge / MOTOR PROTEIN

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