A pertussis toxin-sensitive G protein (GK) links muscarinic cholinergic and A1-purinergic receptors with an inwardly rectifying K+ (KACh) channel in cardiac atrial cell membranes. Although the beta gamma subunits of pertussis toxin-sensitive G proteins (G beta gamma) have been reported to fully activate this channel, it is not known whether exogenously applied G beta gamma interacts with the KACh channel through the same mechanism as the active subunit of endogenous GK (GK*). We examined in inside-out patches the relationship between the concentration of GTP and KACh channel activity with or without preactivation of the channels by either GTP gamma S (guanosine 5'-3-O-(thio)triphosphate) or G beta gamma purified from bovine brain. In the control, KACh channels were activated by intracellular GTP (with acetylcholine in the pipette) in a positive cooperative manner (Hill coefficient approximately 2.5). As the channels were preactivated by GTP gamma S to progressively higher levels, the GTP channel activity relationship shifted more to the left, but the Hill coefficients of the curves remained the same. The same changes were observed when KACh channels were preactivated with brain G beta gamma. These results indicate that endogenous GK* and exogenous G beta gamma share a common molecular mechanism to activate the KACh channel.