Kinetic properties of the ATP-dependent actin-myosin sliding as revealed by the force-movement assay system with a centrifuge microscope.

To study the kinetic properties of the ATP-dependent actin-myosin sliding responsible for muscle contraction, we developed an in vitro force-movement assay system, in which centrifugal forces were applied to myosin-coated polystyrene beads sliding along actin cables of giant algal cells in the presence of ATP. Under constant centrifugal forces directed opposite to the bead movement ("positive" loads), the beads moved with constant velocities. The steady-state force-velocity (P-V) curve thus obtained was double-hyperbolic in shape, being analogous to the P-V curve of single muscle fibers. Under constant centrifugal forces in the direction of the bead movement ("negative" loads), on the other hand, the beads also moved with constant velocities. Unexpectedly, the velocity of bead movement did not increase with increasing negative loads, but decreased markedly (by 20-60%). We also studied the effect of centrifugal forces at right angles with actin cables on the bead movement.