In frog erythrocytes, desensitization of beta-adrenergic receptors is characterized by a decrease in the beta-receptor recognition sites in the plasma membrane and a concomitant increase in the number of this receptor's binding sites in the cytosol. We have documented that this redistribution of the receptor recognition sites reflects the internalization of the surface-bound beta-adrenergic receptors. The present study was addressed to examine whether transglutaminase plays a role in the agonist-mediated internalization of beta-adrenergic receptor recognition sites. Pretreatment of cells with methylamine was found to decrease the internalization and the loss of membrane-bound beta-adrenergic receptors induced by isoproterenol. Methylamine appears to be equally potent in inhibiting transglutaminase activity and in preventing internalization and the receptor loss. The effect of methylamine on soluble and on membrane-bound beta-adrenergic receptors is due to a change in Bmax rather than Kd of these binding sites. Among eight inhibitors of transglutaminase tested, the rank order potency for blocking the enzyme can be correlated with that for preventing the receptor loss and receptor internalization. Moreover, these drug effects on beta-adrenergic receptors are unrelated to the inhibition of isoproterenol-sensitive adenylate cyclase or the binding of [3H]dihydroalprenolol to beta-receptors. These result may lend credence to the view that transglutaminase participates in the internalization and the decrease of membrane-bound receptors during desensitization of beta-adrenergic receptors.