Kinetic parameters of the inhibitory effect of vincristine on Mg2+ATPase activity containing in the actomyosin-like protein from rat sarcoma 45 and in actomyosin from rat skeletal muscle was studied. The alkaloid decreased the VM value for the reaction catalyzed by ATPase in presence of Mg2+ and EGTA and did not alter Km value for both contractile proteins. Inhibitory constants, calculated for the actomyosin from skeletal muscles and for the actomyosin-like protein from sarcoma 45, constituted 520 mcM and 250 mcM, respectively. Vincristine appears to inhibit Mg2+-ATPase containing in these contractile proteins studied by the non-competitive type, simultaneously with lowering of the activating effect of actin on the myosin-derived ATPase.