title: Conformation of the troponin core complex in the thin filaments of skeletal muscle during relaxation and active contraction.
abstract: Contraction of skeletal and cardiac muscles is regulated by Ca(2+) binding to troponin in the actin-containing thin filaments, leading to an azimuthal movement of tropomyosin around the filament that uncovers the myosin binding sites on actin. Here, we use polarized fluorescence to determine the orientation of the C-terminal lobe of troponin C (TnC) in skeletal muscle cells as a step toward elucidating the molecular mechanism of troponin-mediated regulation. Assuming, as shown by X-ray crystallography, that this lobe of TnC is part of a well-defined troponin domain called the IT arm, we show that the coiled coil formed by troponin components I and T makes an angle of about 55° with the thin filament axis in relaxed muscle, in contrast with previous models based on electron microscopy in which this angle is close to 0°. The E helix of TnC makes an angle of about 45° with the thin filament axis. Both the IT coiled coil and the TnC E helix tilt by about 10° on muscle activation. By combining in situ measurements of the orientation of the IT arm and regulatory domain of troponin, which together form the troponin core complex, with published intermolecular distances between thin filament components, we derive models of thin filament structure in which the IT arm of troponin holds its regulatory domain close to the actin surface. Although the structure and function of troponin regions outside the core complex remain to be characterized, the present results provide useful constraints for molecular models of the mechanism of muscle regulation.
authors.affiliation: Randall Division of Cell and Molecular Biophysics and British Heart Foundation Centre of Research Excellence, King's College London, London SE1 1UL, UK.
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Knowles
authors.forename: Malcolm
authors.initials: M
authors.lastname: Irving
authors.forename: Yin-Biao
authors.initials: YB
authors.lastname: Sun
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002645:Chickens; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D009119:Muscle Contraction / Q000502:physiology*; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D009126:Muscle Relaxation; D018482:Muscle, Skeletal / Q000502:physiology*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012518:Sarcomeres / Q000378:metabolism; D019209:Troponin C / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D019209:Troponin C; D002118:Calcium