Filamin binds to F-actin and influences the myosin-actin interaction. At relatively low concentrations, filamin activates actomyosin Mg2+-ATPase, whereas higher concentrations of filamin exert an inhibitory effect. Activation of ATPase activity occurs under conditions where a loose meshwork of actin filaments is present and inhibition is associated with the appearance of closely apposed bundles of actin filaments. Maximum activation (about fourfold) of actomyosin ATPase activity by filamin occurs between 30 and 65 mM KCl, at pH 6.5, and at temperatures not less than 30 degrees C. ATPase activation requires higher concentrations of filamin in the presence than in the absence of tropomyosin. Filamin does not activate Mg2+-ATPase activity of acto-subfragment-1 and has only a slight effect on the Mg2+-ATPase of acto-heavy meromyosin, but it inhibits the activity of both these systems under conditions similar to those that inhibit actomyosin ATPase activity.