The cardiac troponin C mutation Leu29Gln found in a patient with hypertrophic cardiomyopathy does not alter contractile parameters in skinned murine myocardium

摘要

The present study investigates the effects of the first mutation of troponin C (hcTnC^L29Q) found in a patient with hypertrophic cardiomyopathy (HCM) on force–pCa relations and the interplay with phosphorylation of sarcomeric PKA substrates. In triton-skinned murine cardiac fibers, the endogenous mcTnC was extracted and the fibers were subsequently reconstituted with recombinant wild-type and mutant hcTnC. Force–pCa relations of preparations containing hcTnC^L29Q or hcTnC^WT were similar. Incubation of fibers reconstituted with the recombinant proteins with phosphatase to dephosphorylate sarcomeric PKA substrates induced an increase in Ca²⁺ sensitivity, slightly more pronounced (0.04 pCa units) in hcTnC^L29Q-containing fibers. Incubation of the dephosphorylated fibers with PKA induced significant rightward shifts of force–pCa relations of similar magnitude with both, hcTnC^L29Q and hcTnC^WT. No significant effects of hcTnC^L29Q on the velocity of unloaded shortening were observed. In conclusion, no major differences in contractile parameters of preparations containing hcTnC^L29Q compared to hcTnC^WT were observed. Therefore, it appears unlikely that hcTnC^L29Q induces the development of HCM by affecting the regulation of Ca²⁺-activated force and interference with PKA-mediated modulation of the Ca²⁺ sensitivity of contraction.