Single molecule mechanics resolves the earliest events in force generation by cardiac myosin

eLife

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[1] Department of Pathology and Laboratory Medicine, Robert Wood Johnson Medical School, Rutgers University, Piscataway, United States;Graduate Group in Biochemistry and Molecular Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, United States;LENS - European Laboratory for Non-linear Spectroscopy, Sesto Fiorentino, Italy;Department of Physics and Astronomy, University of Florence, Sesto Fiorentino, Italy;Pennsylvania Muscle Institute, Perelman School of Medicine, University of Pennsylvania, Philadelphia, United States;

关键词: myosin;    optical trap;    cardiac myosin;    phosphate release;    working stroke;    strong binding;    Human;   

DOI  :  10.7554/eLife.49266

来源: publisher

【 摘 要 】

10.7554/eLife.49266.001Key steps of cardiac mechanochemistry, including the force-generating working stroke and the release of phosphate (Pi), occur rapidly after myosin-actin attachment. An ultra-high-speed optical trap enabled direct observation of the timing and amplitude of the working stroke, which can occur within <200 μs of actin binding by β-cardiac myosin. The initial actomyosin state can sustain loads of at least 4.5 pN and proceeds directly to the stroke or detaches before releasing ATP hydrolysis products. The rates of these processes depend on the force. The time between binding and stroke is unaffected by 10 mM Pi which, along with other findings, indicates the stroke precedes phosphate release. After Pi release, Pi can rebind enabling reversal of the working stroke. Detecting these rapid events under physiological loads provides definitive indication of the dynamics by which actomyosin converts biochemical energy into mechanical work.

【 授权许可】

CC BY