C-Terminal Basic Region of Troponin T Alters the Ca²⁺-Dependent Changes in Troponin I Interactions

The C-terminal 14-16 residues of human troponin T are required for full inactivation, and they prevent full activation at saturating Ca²⁺. Basic residues within that C-terminal region of TnT are essential for its function, but the mechanism of action is unknown. That region of TnT is natively disordered and does not appear in reconstructions of the troponin structure. We used Förster resonance energy transfer to determine if the C-terminal basic region of TnT alters transitions of TnI or if it operates independently. We also examined Ca²⁺-dependent changes in the C-terminal region of TnT itself. Probes on TnI-143 (inhibitory region) and TnI-159 (switch region) moved away from sites on actin and tropomyosin and toward TnC-84 at high Ca²⁺. Ca²⁺ also displaced C-terminal TnT from actin-tropomyosin but without movement toward TnC. Deletion of C-terminal TnT produced changes in TnI-143 like those effected by Ca²⁺, but effects on TnI-159 were muted; there was no effect on the distance of the switch region to TnC-84. Substituting Ala for basic residues within C-terminal TnT displaced C-terminal TnT from actin-tropomyosin. The results suggest that C-terminal TnT stabilizes tropomyosin in the inactive position on actin. Removal of basic residues from C-terminal TnT produced a Ca²⁺-like state except that the switch region of TnI was not bound to TnC. Addition of Ca²⁺ caused more extreme displacement from actin-tropomyosin as the active state became more fully occupied as in the case of wild-type TnT in the presence of both Ca²⁺ and bound rigor myosin S1.