Contractility parameters of human β-cardiac myosin with the hypertrophic cardiomyopathy mutation R403Q show loss of motor function
| Autor: | Zoltan Ujfalusi, Shirley Sutton, Suman Nag, Stephen J. Langer, Leslie A. Leinwand, Kathleen M. Ruppel, Ruth F. Sommese, Ariana C. Combs, James A. Spudich, Michael A. Geeves |
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| Rok vydání: | 2015 |
| Předmět: |
Myosin light-chain kinase
macromolecular substances 030204 cardiovascular system & hematology Biology Biochemistry Contractility 03 medical and health sciences 0302 clinical medicine maximum entropy method Myosin electron density Research Articles Actin 030304 developmental biology 0303 health sciences Crystallography Multidisciplinary Meromyosin SciAdv r-articles Tropomyosin Troponin Cell biology Metallic alloy High pressure biology.protein MYH7 Chemical bonding Research Article |
| Zdroj: | Science Advances |
| ISSN: | 2375-2548 |
| DOI: | 10.1126/sciadv.1500511 |
| Popis: | Force parameters of human β-cardiac myosin with the hypertrophic cardiomyopathy mutation R403Q show loss of molecular motor function. Hypertrophic cardiomyopathy (HCM) is the most frequently occurring inherited cardiovascular disease. It is caused by mutations in genes encoding the force-generating machinery of the cardiac sarcomere, including human β-cardiac myosin. We present a detailed characterization of the most debated HCM-causing mutation in human β-cardiac myosin, R403Q. Despite numerous studies, most performed with nonhuman or noncardiac myosin, there is no consensus about the mechanism of action of this mutation on the function of the enzyme. We use recombinant human β-cardiac myosin and new methodologies to characterize in vitro contractility parameters of the R403Q myosin compared to wild type. We extend our studies beyond pure actin filaments to include the interaction of myosin with regulated actin filaments containing tropomyosin and troponin. We find that, with pure actin, the intrinsic force generated by R403Q is ~15% lower than that generated by wild type. The unloaded velocity is, however, ~10% higher for R403Q myosin, resulting in a load-dependent velocity curve that has the characteristics of lower contractility at higher external loads compared to wild type. With regulated actin filaments, there is no increase in the unloaded velocity and the contractility of the R403Q myosin is lower than that of wild type at all loads. Unlike that with pure actin, the actin-activated adenosine triphosphatase activity for R403Q myosin with Ca2+-regulated actin filaments is ~30% lower than that for wild type, predicting a lower unloaded duty ratio of the motor. Overall, the contractility parameters studied fit with a loss of human β-cardiac myosin contractility as a result of the R403Q mutation. |
| Databáze: | OpenAIRE |
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