Ensemble Force Changes that Result from Human Cardiac Myosin Mutations and a Small-Molecule Effector

• DOI: 10.1016/j.celrep.2015.04.006
• Publication Date: 2015-05-02T22:40:45Z
• Authors: Tural Aksel; Elizabeth Choe Yu; Shirley Sutton; Kathleen M. Ruppel; James A. Spudich
• Keywords: Adenosine Triphosphatases; Models, Molecular; 0303 health sciences; Utrophin; QH301-705.5; Statistics as Topic; Models, Biological; Article; Actins; Biomechanical Phenomena; 3. Good health; Small Molecule Libraries; Ventricular Myosins; Actin Cytoskeleton; Mice; 03 medical and health sciences; Mutation; Animals; Humans; Urea; Cattle; Biology (General); Cardiomyopathies; Software

Cardiomyopathies due to mutations in human β-cardiac myosin are a significant cause of heart failure, sudden death, and arrhythmia. To understand the underlying molecular basis of changes in the contractile system's force production due to such mutations and search for potential drugs that restore force generation, an in vitro assay is necessary to evaluate cardiac myosin's ensemble force using purified proteins. Here, we characterize the ensemble force of human α- and β-cardiac myosin isoforms and those of β-cardiac myosins carrying left ventricular non-compaction (M531R) and dilated cardiomyopathy (S532P) mutations using a utrophin-based loaded in vitro motility assay and new filament-tracking software. Our results show that human α- and β-cardiac myosin, as well as the mutants, show opposite mechanical and enzymatic phenotypes with respect to each other. We also show that omecamtiv mecarbil, a previously discovered cardiac-specific myosin activator, increases β-cardiac myosin force generation.