Effect of the state of oxidation of cysteine 190 of tropomyosin on the assembly of the actin-tropomyosin complex.
Tropomyosin, cross-linked at cysteine 190, was found to bind more weakly to actin filaments than uncross-linked tropomyosin. Cross-linking of tropomyosin can cause actin filaments nearly completely covered with tropomyosin to be uncovered almost completely. The critical monomer concentration of actin is not significantly changed by binding of cross-linked or uncross-linked tropomyosin to actin filaments. The binding curves were analyzed quantitatively, thereby taking into account the polar end-to-end contact of tropomyosin molecules bound by actin and the overlap of the seven subunit binding sites along the actin filament. Under the conditions of the experiment (80 mM KCl, 1 mM MgCl2, pH 7.5, 38-42 degrees C), the equilibrium constant for isolated binding of tropomyosin to actin filaments is in the range 1 x 10(3)-3 x 10(3) M-1. The equilibrium constants for binding of tropomyosin to binding sites along the actin filament with one or two neighbouring tropomyosin molecules are in the range of 10(6) or 10(8) to 10(9) M-1, respectively. The equilibrium constants for binding of tropomyosin to binding sites along the actin filament with one or two neighbouring tropomyosin molecules are in the range of 10(6) or 10(8) to 10(9) M-1, respectively. The equilibrium constants for cross-linked and uncross-linked tropomyosin differ by a factor of only about two. Owing to the highly cooperative binding, these differences are sufficient so that actin filaments nearly completely covered with uncross-linked tropomyosin are uncovered almost completely by cross-linking tropomyosin at cysteine 190.
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D003545:Cysteine; D007700:Kinetics; D008027:Light; D046911:Macromolecular Substances; D010084:Oxidation-Reduction; D011485:Protein Binding; D012542:Scattering, Radiation; D014335:Tropomyosin / Q000378:metabolism*
D000199:Actins; D046911:Macromolecular Substances; D014335:Tropomyosin; D003545:Cysteine