Switch between Large Hand-Over-Hand and Small Inchworm-like Steps in Myosin VI

Many biological motor molecules move within cells using stepsizes predictable from their structures. Myosin VI, however, has much larger and more broadly distributed stepsizes than those predicted from its short lever arms. We explain the discrepancy by monitoring Qdots and gold nanoparticles attached to the myosin-VI motor domains using high-sensitivity nanoimaging. The large stepsizes were attributed to an extended and relatively rigid lever arm; their variability to two stepsizes, one large (72 nm) and one small (44 nm). These results suggest that there exist two tilt angles during myosin-VI stepping, which correspond to the pre- and postpowerstroke states and regulate the leading head. The large steps are consistent with the previously reported hand-over-hand mechanism, while the small steps follow an inchworm-like mechanism and increase in frequency with ADP. Switching between these two mechanisms in a strain-sensitive, ADP-dependent manner allows myosin VI to fulfill its multiple cellular tasks including vesicle transport and membrane anchoring. [Display omitted] ► Brownian search and stepsizes of myosin VI heads were precisely measured ► The broad distribution of stepsizes was due to a mixture of large and small stepsizes ► Large stepsizes were caused by an extended, relatively rigid lever arm ► Myosin VI switches between large and small steps in an ADP dependent manner