Thin Filament Interaction and Ca2+-Desensitization Effect of the C-Terminal End Peptide of Cardiac Troponin T Where Loss of Function Mutations Cause Hypertrophic Cardiomyopathy

BackgroundTroponin T (TnT) is the tropomyosin (Tm)-binding subunit of troponin with a central role in regulating cardiac muscle contractility. The recently identified Tm-binding site 3 in the highly conserved C-terminal end segment of TnT has a troponin I (TnI)-like inhibitory function. Conformational modulation by proteolytic removal of the N-terminal variable region of cardiac TnT (cTnT-ND) in adaptation to inotropy-afterload mismatch enhances the function of Tm-binding site 3 to adjust ventricular contractile kinetics and sustain stroke volume. Mutations in this segment of cTnT cause overactivation of cardiac muscle and hypertrophic cardiomyopathy (HCM). MethodsThe C-terminal end 14 amino acid peptide of cTnT (cTnT-C14) was analyzed for interactions with Tm, F-actin, and F-actin-Tm thin filament using localized surface plasmon resonance (LSPR). Wild-type and HCM mutant cTnT-C14 peptides were used to treat skinned cardiac muscle strips from wild-type and cTnT-ND transgenic mice to assess the effect on Ca{superscript 2}- activation of contraction. ResultscTnT-C14 peptide showed saturable binding to Tm and F-actin in LSPR with physiological affinity, which is significantly impaired by HCM mutation R278C, K280N or R286C. Treatment of wild-type mouse cardiac muscle strips with cTnT-C14 peptide produced a Ca2+- desensitization effect on myofilament activation, which was not seen with cTnT-ND mouse cardiac muscle strips implicating a prior utilization of the same mechanism. The HCM mutant peptides lost this function. ConclusionsThe findings demonstrate the function of the cTnT C-terminal end segment underlying the pathophysiology of the HCM mutations. Its preserved functionality in the form of free peptide presents a drug candidate for the treatment of heart failure.