Truncated myoV-4IQ showed an increased frequency of azimuthal steps compared to native-length myoV-6IQ, but the magnitude of these deflections was independent of lever-arm length.
The azimuthal deflections of myosin V on actin are more common for the truncated lever arm, but the range of these deflections is relatively independent of its lever-arm length.
Myosin V (myoV) is a two-headed myosin capable of taking many successive steps along actin per diffusional encounter, enabling it to transport vesicular and ribonucleoprotein cargos in the dense and complex environment within cells. To better understand how myoV navigates along actin, we used polarized total internal reflection fluorescence microscopy to examine angular changes of bifunctional rhodamine probes on the lever arms of single myoV molecules in vitro. With a newly developed analysis technique, the rotational motions of the lever arm and the local orientation of each probe relative to the lever arm were estimated from the probe's measured orientation. This type of analysis could be applied to similar studies on other motor proteins, as well as other proteins with domains that undergo significant rotational motions. The experiments were performed on recombinant constructs of myoV that had either the native-length (six IQ motifs and calmodulins CaMs) or truncated (four IQ motifs and CaMs) lever arms. Native-length myoV-6IQ mainly took straight steps along actin, with occasional small azimuthal tilts around the actin filament. Truncated myoV-4IQ showed an increased frequency of azimuthal steps, but the magnitudes of these steps were nearly identical to those of myoV-6IQ. The results show that the azimuthal deflections of myoV on actin are more common for the truncated lever arm, but the range of these deflections is relatively independent of its lever-arm length.
Lewis et al. (Mon,) reported a other. Truncated lever arm (myoV-4IQ) vs. Native-length lever arm (myoV-6IQ) was evaluated on Azimuthal deflections on actin. Truncated myoV-4IQ showed an increased frequency of azimuthal steps compared to native-length myoV-6IQ, but the magnitude of these deflections was independent of lever-arm length.