Cardiac troponin C (cTnC) is the calcium (Ca2+) sensing component of the troponin complex. Binding of Ca2+ to cTnC triggers a cascade of myofilament conformational changes that culminate in force production. Mutations in cTnC linked to hypertrophic myocardial myopathy (HCM) induce a greater degree and duration of Ca2+ binding, which may underly the hypertrophic phenotype. Recent evidence from our laboratories demonstrated novel modifications of cTnC Ca2+ binding by cellular magnesium (Mg2+) that we hypothesize may be of significance in promoting HCM.Regulation of contraction has long been thought to occur exclusively through Ca2+ binding to site II of cTnC. However, abundant cellular Mg2+ is a potential competitor for binding to the same sites; work by several groups also suggests this is possible. We have used isothermal titration calorimetry (ITC) to explore the thermodynamic properties associated with the interaction between Ca2+/Mg2+ and site II of cTnC; these experiments demonstrated that physiological concentrations of Mg2+ may compete with Ca2+ to bind site II of cTnC.In experiments reported here, we studied a series of mutations in cTnC thought to be causal in HCM. Three mutants (A8V, L29Q, and A31S) slightly elevated the affinity for both Ca2+ and Mg2+, whereas other mutants (L48Q, Q50R, and C84Y), that are closer to the C-terminal domain and surrounding the EF hand binding motif of site II had a more significant effect on affinity and the thermodynamics of the binding interaction.
Mutation of the high affinity calcium binding sites in cardiac troponin C.
