Effects of G protein beta gamma subunits from rat brain on cardiac K+ channel was examined in single atrial cells of guinea-pig, using patch clamp techniques. We found that 10 pM concentration of rat brain beta gamma subunits preparation could activate the atrial muscarine receptor-gated K+ channel (IK.ACh). Neither the detergent, CHAPS, used to suspend beta gamma nor the boiled beta gamma preparation activated IK.ACh. Furthermore, preincubation of beta gamma subunits preparation in Mg2+-free solution, which easily inactivated alpha-GTP-gamma S, did not affect beta gamma-activation of IK.ACh. We concluded, therefore, that beta gamma subunits themselves can activate IK.ACh.