The effect of guanine nucleotides on the binding of [3H] Quinuclidinyl benzilate to heart muscarinic receptors was studied. It was observed that GTP and Gpp (NH)p increased antagonist binding. Scatchard transformation of the data resulted in curvilinear plots (downward concavity) consistent with positive cooperativity. Graphic analysis for cooperativity indicated that guanine nucleotides: a) increased slightly but consistently the affinity for the antagonist (independent of receptor occupancy) and b) induce positive cooperativity in the binding of [3H]QNB (dependent of receptor occupancy).