EFFECT OF LATERAL FORCES ON THE MOVEMENT OF MYOSIN-COATED BEADS ON ACTIN CABLES STUDIED USING A CENTRIFUGE MICROSCOPE

We developed an in vitro motility assay system, in which myosin-coated polystyrene beads were made to slide on actin filament arrays (actin cables) in giant algal cells and subjected to centrifugal forces, whic h were parallel to the direction of bead movement to serve as external loads on actin-myosin sliding (Oiwa et al. (1990) Proc Natl Acad Sci USA 87: 7893-7897), and succeeded in determining the steady-state forc e-velocity relation of ATP-dependent actin-myosin sliding. To give fur ther information about the properties of actin-myosin sliding, we have applied centrifugal forces, in parallel with the plane of actin-myosi n sliding but at right angles with the direction of bead movement, and have found that such ''lateral'' centrifugal forces reduced the veloc ity of bead movement. In addition, we have also found that the velocit y of bead movement is reduced more markedly with lateral forces applie d from the left side of the bead (''left'' lateral forces) than those applied form the right side of the bead (''right'' lateral forces). Th ese results are discussed in connection with the direction of sliding force generated by the myosin heads on the bead which interact with th e right-handed double helix of actin monomers constituting actin filam ents.