Cellular G protein-coupled receptor kinase levels regulate sensitivity of the {alpha}2b-adrenergic receptor to undergo agonist-induced down-regulation.

Chronic coactivation of alpha(2B)- and beta(2)-adrenoceptors (AR) was recently reported to down-regulate the alpha(2B)-AR at a lower threshold epinephrine (EPI) concentration compared with the activation of alpha(2B)-AR alone. This is the result of a modest beta(2)-AR-dependent up-regulation of G protein-coupled receptor kinase 3 (GRK3). In the present study, we determined that increasing GRK2 or GRK3 levels, independent of beta(2)-AR activation, decreases the EC(50) concentration for agonist-induced down-regulation of the alpha(2B)-AR using NG108 cells with or without overexpression (2- to 10-fold) of GRK2 or GRK3. In parental NG108 cells, the EC(50) concentration of EPI required for down-regulation of the alpha(2B)-AR is 30 microM. A 2- to 3-fold overexpression of GRK3 in NG108 cells, however, reduces the EC(50) to 0.2 microM (a 150-fold decrease), whereas a comparable overexpression of GRK2 reduces it to 1 microM (a 30-fold decrease). However, when GRK3 or GRK2 in NG108 cells are overexpressed 8- to 10-fold, the EC(50) concentration (0.02 microM EPI) for alpha(2B)-AR down-regulation is reduced 1000-fold. These data clearly suggest that a modest (2- to 3-fold) up-regulation of GRK3 is more effective at enhancing the sensitivity of alpha(2B)-AR to down-regulation after exposure to EPI than a modest up-regulation of GRK2, but that both GRK2 and GRK3 are equally effective at inducing alpha(2B)-AR down-regulation when up-regulated 8- to 10-fold. To our knowledge, this is the first report to systematically demonstrate that GRKs, particularly GRK3, play a pivotal role in modulating the agonist EC(50) concentration that down-regulates the alpha(2B)-AR and thus adds a new dimension to an already intricate signaling network.