Tropomyosin (Tm) consists of 7 quasiequivalent repeats known as “periods,” and its specific function may be associated with these periods. To test the hypothesis that either period 2 or 3 promotes force generation by inducing a positive allosteric effect on actin, we reconstituted the thin filament with mutant Tm in which either period 2 (Δ2Tm) or period 3 (Δ3Tm) was deleted. We then studied: isometric tension, stiffness, 6 kinetic constants, and the pCa-tension relationship. N-terminal acetylation of Tm did not cause any differences. The isometric tension in Δ2Tm remained unchanged, and was reduced to ~60% in Δ3Tm. Although the kinetic constants underwent small changes, the occupancy of strongly attached cross-bridges was not much different. The Hill factor (cooperativity) did not differ significantly between Δ2Tm (1.79 ± 0.19) and the control (1.73 ± 0.21), or Δ3Tm (1.35 ± 0.22) and the control. In contrast, pCa50 decreased slightly in Δ2Tm (5.11 ± 0.07), and increased significantly in Δ3Tm (5.57 ± 0.09) compared to the control (5.28 ± 0.04). These results demonstrate that, when ions are present at physiological concentrations in the muscle fiber system, period 3 (but not period 2) is essential for the positive allosteric effect that enhances the interaction between actin and myosin, and increases isometric force of each cross-bridge.
Temperature effect on isometric tension is mediated by regulatory proteins tropomyosin and troponin in bovine myocardium
