Myosins are a superfamily of motor proteins that drive many intracellular processes through transducing chemical energy from ATP hydrolysis into mechanical work. The key event in the transduction process is the myosin “power stroke”, a 60–70° rotation of a long alpha helical coil that is believed to be coupled to the release of Pi following ATP hydrolysis, creating a cycle of attachment, pulling, and detachment for myosin-actin. Earlier models depicted myosin Pi release and power stroke as tightly coupled events. However, later experiments showed that Pi release and power stroke occur at different times and rates, making their sequence unresolved. We used a single-headed myosin Va construct containing a (S217A) mutation, that slows Pi release from the active site, in a laser trap assay to determine this sequence of events. The myosin Va construct was used to see the effect of Pi and resistive load on the size of the primary power stroke, secondary power stroke (hitch), and the duration of the actomyosin binding event (ton). The power stroke and the hitch were unaffected at every resistive load at 0 mM Pi, but the hitch and ton were significantly reduced by 30 mM Pi at the highest resistive load. Thus, these observations suggest that the power stroke occurs while Pi remains in myosin’s active site, implying that Pi release follows the power stroke.