Propafenone, a class I antiarrhythmic agent, inhibits several membrane currents (I(Na), I(Ca), I(K), Ito), however, its effects on ATP-sensitive potassium current (I(K)ATP) of cardiac cells have not been tested. We evaluated the blocking effects of 0.1 to 100 microM propafenone applications at 35 degrees C on the whole-cell I(K)ATP as triggered by dinitrophenol (75 microM) in adult rabbit dissociated atrial and ventricular cardiomyocytes in comparison. The block of I(K)ATP by propafenone was dose-dependent, fully reversible and voltage-independent. The dose-response relation, as evaluated at 0 mV for atrial myocytes (ED50 = 1.26+/-0.17 microM, Hill number = 1.25+/-0.22) was significantly shifted to the left vs. that in ventricular myocytes (ED50 = 4.94+/-0.59 microM, Hill number = 1.22+/-0.14). It is concluded that propafenone blocks cardiac I(K)ATP at a single site with 4 times higher affinity for the drug in atrial myocytes. This block of cardiac I(K)ATP might play a role in the beneficial and adverse effects of the drug.