Mutations in the MYBPC3 gene, encoding cardiac myosin-binding protein C (cMyBP-C), are a frequent cause of familial hypertrophic cardiomyopathy. In the present study, we investigated whether protein composition and function of the sarcomere are altered in a homogeneous familial hypertrophic cardiomyopathy patient group with frameshift mutations in MYBPC3 (MYBPC3mut).Methods and Results- Comparisons were made between cardiac samples from MYBPC3 mutant carriers (c.2373dupG, n=7; c.2864-2865delCT, n=4) and nonfailing donors (n= 13). Western blots with the use of antibodies directed against cMyBP-C did not reveal truncated cMyBP-C in MYBPC3mut. Protein expression of cMyBP-C was significantly reduced in MYBPC3mutby 33 ±5%. Cardiac MyBP-C phosphorylation in MYBPC3mutsamples was similar to the values in donor samples, whereas the phosphorylation status of cardiac troponin I was reduced by 84 ±5%, indicating divergent phosphorylation of the 2 main contractile target proteins of the β-adrenergic pathway. Force measurements in mechanically isolated Triton-permeabilized cardiomyocytes demonstrated a decrease in maximal force per cross- sectional area of the myocytes in MYBPC3mut(20.2±2.7 kN/m2) compared with donor (34.5± 1.1 kN/m2). Moreover, Ca2+sensitivity was higher in MYBPC3mut(pCa50=5.62±0.04) than in donor (pCa50=5.54±0.02), consistent with reduced cardiac troponin I phosphorylation. Treatment with exogenous protein kinase A, to mimic β-adrenergic stimulation, did not correct reduced maximal force but abolished the initial difference in Ca sensitivity between MYBPC3mut(pCa50=5.46±0.03) and donor (pCa50=5.48±0. 02).Conclusions- Frameshift MYBPC3 mutations cause haploinsufficiency, deranged phosphorylation of contractile proteins, and reduced maximal force-generating capacity of cardiomyocytes. The enhanced Ca2+sensitivity in MYBPC3mutis due to hypophosphorylation of troponin I secondary to mutation-induced dysfunction.

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Circulation (Baltimore)
Erasmus MC: University Medical Center Rotterdam

van Dijk, S.J, Dooijes, D, Remedios, C.D, Michels, M, Lamers, J.M.J, Winegrad, S, … van Velden, J.D. (2009). Cardiac myosin-binding protein C mutations and hypertrophic ardiomyopathy haploinsufficiency, deranged phosphorylation, and cardiomyocyte dysfunction. Circulation (Baltimore), 119(11), 1473–1483. doi:10.1161/CIRCULATIONAHA.108.838672