Solution NMR Structure of the Junction between Tropomyosin Molecules: Implications for Actin Binding and Regulation

• Published in: Journal of molecular biology 2006-11, Vol.364 (1), p.80-96
• Main Authors: Greenfield, Norma J., Huang, Yuanpeng Janet, Swapna, G.V.T., Bhattacharya, Aneerban, Rapp, Brian, Singh, Abhishek, Montelione, Gaetano T., Hitchcock-DeGregori, Sarah E.
• Format: Article
• Language: English
• Subjects: actin binding site; Actins - metabolism; Amino Acid Sequence; Animals; coiled coil; conformational flexibility; Crystallography, X-Ray; four-helix bundle; Models, Molecular; Molecular Sequence Data; Multiprotein Complexes; Nuclear Magnetic Resonance, Biomolecular; Peptides - chemistry; Peptides - genetics; Peptides - metabolism; Protein Binding; Protein Structure, Secondary; Protein Structure, Tertiary; Rats; Tropomyosin - chemistry; Tropomyosin - metabolism; tropomyosin overlap complex
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2006.08.033

Tropomyosin is a coiled-coil protein that binds head-to-tail along the length of actin filaments in eukaryotic cells, stabilizing them and providing protection from severing proteins. Tropomyosin cooperatively regulates actin's interaction with myosin and mediates the Ca 2+-dependent regulation of contraction by troponin in striated muscles. The N-terminal and C-terminal ends are critical functional determinants that form an “overlap complex”. Here we report the solution NMR structure of an overlap complex formed of model peptides. In the complex, the chains of the C-terminal coiled coil spread apart to allow insertion of 11 residues of the N-terminal coiled coil into the resulting cleft. The plane of the N-terminal coiled coil is rotated 90° relative to the plane of the C terminus. A consequence of the geometry is that the orientation of postulated periodic actin binding sites on the coiled-coil surface is retained from one molecule to the next along the actin filament when the overlap complex is modeled into the X-ray structure of tropomyosin determined at 7 Å. Nuclear relaxation NMR data reveal flexibility of the junction, which may function to optimize binding along the helical actin filament and to allow mobility of tropomyosin on the filament surface as it switches between regulatory states.