To determine the palmitoylation sites in the human dopamine D(1) receptor, we expressed wild type and mutant receptors in which candidate cysteines in the carboxyl tail were substituted by alanines both individually (A347, A351) and together (AA). Our results showed that palmitoylation levels of A347 and A351 were reduced substantially and that AA had no detectable signal of palmitoylation. These data indicate that cysteines 347 and 351 were both palmitoylated and that they were the only sites of palmitoylation. We introduced a cAMP-dependent protein kinase site encompassing the position 351. We predicted that a functional cAMP-dependent protein kinase site would impair receptor-G protein coupling if it is not occluded by palmitoylation. Our results demonstrated that indeed, the introduction of the cAMP-dependent protein kinase site caused reduced potency of dopamine stimulation of adenylyl cyclase, and thus confirmed that when unoccluded, the cAMP-dependent protein kinase site introduced to position 351 of dopamine D(1) receptor could confer constitutive desensitization.