Interplay between the overlapping ends of tropomyosin and the N terminus of cardiac troponin T affects tropomyosin states on actin.

The functional significance of the molecular swivel at the head-to-tail overlapping ends of contiguous tropomyosin (Tm) dimers in striated muscle is unknown. Contractile measurements were made in muscle fibers from transgenic (TG) mouse hearts that expressed a mutant α-Tm (Tm(H276N)). We also reconstituted mouse cardiac troponin T (McTnT) N-terminal deletion mutants, McTnT(1-44Δ) and McTnT(45-74Δ), into muscle fibers from Tm(H276N). For controls, we used the wild-type (WT) McTnT because altered effects could be correlated with the mutant forms of McTnT. Tm(H276N) slowed crossbridge (XB) detachment rate (g) by 19%. McTnT(1-44Δ) attenuated Ca(2+)-activated maximal tension against Tm(WT) (36%) and Tm(H276N) (38%), but sped g only against Tm(H276N) by 35%. The rate of tension redevelopment decreased (17%) only in McTnT(1-44Δ) + Tm(H276N) fibers. McTnT(45-74Δ) attenuated tension (19%) and myofilament Ca(2+) sensitivity (pCa50=5.93 vs. 6.00 in the control fibers) against Tm(H276N), but not against Tm(WT) background. Thus, altered XB cycling kinetics decreased the fraction of strongly bound XBs in McTnT(1-44Δ) + Tm(H276N) fibers, whereas diminished thin-filament cooperativity attenuated tension in McTnT(45-74Δ) + Tm(H276N) fibers. In summary, our study is the first to show that the interplay between the N terminus of cTnT and the overlapping ends of contiguous Tm effectuates different states of Tm on the actin filament.