Mutated human beta 3-adrenergic receptor (Trp64Arg) lowers the response tobeta 3-adrenergic agonists in transfected 3T3-L1 preadipocytes

Wild-type or mutated human beta 3-adrenergic receptor (Trp64Arg) cDNAs were stably expressed in mouse 3T3-L1 cells. Saturation binding study using a b eta-adrenergic ligand revealed that there was no significant difference in the receptor density and the equilibrium dissociation constant between the two cell lines. However, the ability of the mutant beta 3-adrenergic recept or to accumulate cyclic AMP (cAMP) in response to isoproterenol was much re duced and Kact for cAMP accumulation was lowered as compared to the wild ty pe receptor. The amount of alpha subunit of stimulatory GTP-binding protein (GS alpha) and adenylyl cyclase activity in response to forskolin were not different in the two cell lines. The responses of the mutant receptor to e pinephrine, norepinephrine and L-755,507, a highly specific agonist for hum an beta 3-adrenergic receptor, were also reduced, but the reduction of Kact for L-755,507 was more evident than other agonists tested. The cAMP accumu lation in response to some conventional beta 3 agonists was less than 10% o f that to isoproterenol even in the cells expressing the wild type receptor . These results suggest that the Trp64Arg mutant beta 3-adrenergic receptor has less ability to stimulate adenylyl cyclase, and that lipolytic activit y through the beta 3-adrenergic receptor by catecholamines in subjects carr ying this mutation might be suppressed.