Ser165 of transmembrane helix IV is not involved in the interaction of catecholamines with the alpha-2a-adrenoceptor.

Molecular modeling studies have predicted that the β-hydroxyl group of the catecholamines interacts with the β2-adrenoceptor at the serine residue at position 165 (Ser165) located on transmembrane helix IV; however, this has not been confirmed by site-directed mutagenesis. It has been inferred that this site, which is conserved in all of the nine known α- and β-adrenoceptor subtypes, is also involved in the interaction of catecholamines with the α2a-adrenoceptor. To test the hypothesis that the β-hydroxyl group of the catecholamines interacts with Ser165 of the α2a-adrenoceptor, we prepared a mutant α2a-adrenoceptor where Ser165 was mutated to alanine. Mutation of Ser165 of the cα2a-adrenoceptor to alanine had no effect on the affinity of dopamine (which lacks the β-hydroxyl group) or either enantiomer of norepinephrine or epinephrine (both of which possess the β-hydroxyl group), indicating that Ser165 is not involved in the interaction of the catecholamines with the α2a-adrenoceptor. We have previously shown that mutation of Ser90, located in transmembrane helix II, to either alanine or cysteine produces a selective reduction in the affinity of the (–)-enantiomers of the catecholamines for the α2a-adrenoceptor, with no effect on the (+)-enantiomers or the corresponding β-desoxy analogs. This is consistent with the known stereoselectivity involved in the interactions of catecholamines with the α2a-adrenoceptor. The results of the present investigation indicate that Ser165 is not involved in the interaction of catecholamines with the α2a-adrenoceptor. Because all known α-adrenoceptor subtypes have a serine residue at a position corresponding to Ser90 of the α2a-adrenoceptor, it would appear that this site represents an important point for attachment of the β-hydroxyl group of catecholamines.