Aconiti tuber butanol fraction has been recently known to have stimulatory effect on myocardial contractility. In the present study, the possibility that the Aconiti tuber butanol fraction acts directly on contractile proteins of myocardium has been investigated using natural actomyosin extracted from dog heart. It revealed that Aconiti tuber butanol fraction in concentrations from 10-2 ~ 10-7 gm/ml had no stimulatory effect on either the Mg++ or Ca++-activated adenosinetriphosphatase activity of cardiac actomyosin. And no direct Ca++-like action of the drug on cardiac actomyosin was also found. Aconiti tuber butanol fraction in concentrations above 10-4 gm/ml, however, was somewhat stimulatory on superprecipitation of actomyosin and markedly inhibited the membrane bound Na+-K+-activated ATPase activity. In these connections, the positive inotropic action of Aconiti tuber butanol fraction on myocardium thus does not seem to reflect a direct interaction with contractile proteins, but the drug seem to stimulate myocardial contractility through the actions on the membrane transport of Ca++.