MYK-461 reduces the activity of homeostasis ATPASE by inhibiting the phospharic acid release rate of β -myocardial myosin -S1. MYK-461 regulates several steps in the myosin chemo-mechanical cycle: in addition to reducing the rate-limiting step (phospharic acid release) in this cycle, MYK-461 also reduces the number of myosin-S1 heads that interact with actin filaments during the transition from weak to strong binding states. MYK-461 also reduced the binding rate of myosin to ADP-actin and the release rate of ADP from Myosin-S1.

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