Caldesmon tethers myosin V to actin and facilitates in vitro motility.

The current study examines the hypothesis that caldesmon can facilitate the interaction of myosin V with actin filaments by tethering myosin V to actin. Myosin V, purified from bovine brain stem, translocated actin filaments in an in vitro motility assay at a velocity of 0.30+/-0.05 microm/s in the absence of caldesmon at a myosin concentration of 50 microg/ml (ionic strength=110 mM). Filament binding and motility was absent when the myosin concentration applied to the coverslip was reduced to 5 microg/ml, however, the addition of 0.4 microM caldesmon restored binding and motility (0.28+/-0.06 microm/s). This restoration of motility in the presence of caldesmon was blocked by an N-terminal fragment of caldesmon that competitively inhibits the binding of intact caldesmon to myosin. Similar to previous findings with both smooth muscle myosin and platelets (Haeberle et al., 1992; Hemric et al., 1994), these results demonstrate that caldesmon can form a mobile tether that maintains the proximity of myosin V with actin filaments without restricting filament sliding.