Unphosphorylated myosin acts as a load to slow down the rate at which actin is moved by faster cycling phosphorylated cross-bridges in an in vitro motility assay.
Does the ratio of unphosphorylated to phosphorylated myosin modulate the rate of actin filament sliding in vitro?
Unphosphorylated myosin acts as a mechanical load that slows down actin filament sliding driven by phosphorylated myosin cross-bridges in vitro.
Although it is generally believed that phosphorylation of the regulatory light chain of myosin is required before smooth muscle can develop force, it is not known if the overall degree of phosphorylation can also modulate the rate at which cross-bridges cycle. To address this question, an in vitro motility assay was used to observe the motion of single actin filaments interacting with smooth muscle myosin copolymers composed of varying ratios of phosphorylated and unphosphorylated myosin. The results suggest that unphosphorylated myosin acts as a load to slow down the rate at which actin is moved by the faster cycling phosphorylated cross-bridges. Myosin that was chemically modified to generate a noncycling analogue of the "weakly" bound conformation was similarly able to slow down phosphorylated myosin. The observed modulation of actin velocity as a function of copolymer composition can be accounted for by a model based on mechanical interactions between cross-bridges.
Warshaw et al. (1990) studied this question. Varying ratios of phosphorylated and unphosphorylated myosin was evaluated on Actin filament sliding velocity. Unphosphorylated myosin acts as a load to slow down the rate at which actin is moved by faster cycling phosphorylated cross-bridges in an in vitro motility assay.