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

• Main Authors: Michael S Woody, Donald A Winkelmann, Marco Capitanio, E Michael Ostap, Yale E Goldman
• Format: Article
• Language: English
• Published: eLife Sciences Publications Ltd 2019-09-01
• Series: eLife
• Subjects: myosin; optical trap; cardiac myosin; phosphate release; working stroke; strong binding
• Online Access: https://elifesciences.org/articles/49266

Key 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.