Troponin I inhibitory peptide suppresses the force generation in smooth muscle by directly interfering with cross-bridge formation.

To explore possible mechanisms involving the thin filament-linked regulation of contraction in living smooth muscles, we studied the effects of a synthetic peptide of rabbit cardiac troponin I [residues 136-147] (TnIp), which is a minimal sequence required to inhibit striated muscle acto-tropomyosin-myosin ATPase activity, on the mechanical properties of beta-escin skinned preparations of taenia caeci from guinea pig. TnIp reversibly suppressed the Ca(2+)-activated force without significant effects on the Ca(2+) sensitivity and on the phosphorylation level of myosin regulatory light chain (MLC(20)). TnIp also reversibly suppressed the Ca(2+)/calmodulin-independent contraction induced by 30mM Mg(2+). An analogue of TnIp, which lost inhibiting action on acto-tropomyosin-myosin ATPase activity, affected neither Ca(2+)-activated nor 30mM Mg(2+)-induced contraction. These results indicate that TnIp suppresses the force generation in smooth muscle by directly interfering with cross-bridge formation rather than inhibiting the Ca(2+)/calmodulin-dependent thick and thin filament activating processes.