The role of the guanine nucleotide exchange reaction in the regulation of the beta-adrenergic receptor and in the actions of catecholamines and cholera toxin on adenylate cyclase in turkey erythrocyte membranes.

Several changes were noted in the characteristics of the turkey erythrocyte beta-adrenergic receptor and in the kinetic properties of adenylate cyclase following pretreatment of erythrocyte membranes with isoproterenol and GMP, and thorough washing to remove these agents. The changes include modifications in the binding of agonist (isoproterenol) and in revelation of marked effects of GTP on agonist binding; reduction in the lag in Gpp(NH)p activation of adenylate cyclase; short lived activation by GTP which is lengthened by treatment with cholera toxin and NAD prior to pretreatment with isoproterenol and GMP. Treatment with cholera toxin also shortened the lag in activation by Gpp(NH)p and increased the steady state levels of activation by both Gpp(NH)p and GTP. The following conclusions can be drawn: (i) catecholamines, in the presence of a guanine nucleotide, stimulate the exchange of bound and exogenous nucleotide; (ii) the exchange reaction is involved in both the activation of adenylate cyclase and in the reciprocal effects of hormone and guanine nucleotides on each other's binding: (iii) the beta-adrenergic receptor and nucleotide regulatory components are linked in turkey erythrocyte membranes; (iv) both cholera toxin and catecholamines, although by different mechanisms, stimulate the exchange reaction at the nucleotide regulatory sites.