Desensitization, phosphorylation and palmitoylation of the human dopamine D1 receptor

Eur J Pharmacol. 1994 Mar 15;267(1):7-19. doi: 10.1016/0922-4106(94)90219-4.


The regulation and post-translational modifications of the human dopamine D1 receptor were studied in the baculovirus-eukaryotic cell expression system. Baculovirus constructs containing either the DNA encoding the dopamine D1 receptor or a DNA encoding a c-myc epitope tagged dopamine D1 receptor (c-myc-dopamine D1 receptor) were used to infect Spodoptera frugiperda (Sf9) insect cells. Expressed dopamine D1 and c-myc-dopamine D1 receptors bound agonists and antagonists with affinities and a rank order of potency characteristic of a classical dopamine D1 receptor pharmacological profile. In membrane preparations from cells expressing c-myc-dopamine D1 receptor, the photoaffinity label [125I](3-methyl-2-[4'-azidophenyl]-2,3,5-tetrahydro-2H-3-benzazepine) ([125I]MAB) bound specifically upon photolysis. A major broad band of approximately 48 kDa was detected. This species was identified in immunoblots by the monoclonal antibody raised against the c-myc epitope of c-myc-dopamine D1 receptor was isolated by immunoprecipitation from whole cells and was shown to be post-translationally modified by phosphorylation and palmitoylation. Exposure of cells expressing c-myc-dopamine D1 receptor to dopamine for 15 min resulted in a reduction in the maximal dopamine stimulated adenylyl cyclase activity, which was accompanied by an increased phosphorylation of the receptor and a rapid redistribution of surface c-myc-dopamine D1 receptor as detected by in situ immunofluorescence. Dopamine exposure also resulted in an increased level of incorporation of [3H]palmitic acid into the receptor. Thus, we provide the first evidence that the human dopamine D1 receptor undergoes agonist-dependent desensitization, phosphorylation and palmitoylation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine / metabolism
  • 2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine / pharmacology
  • Adenylyl Cyclases / analysis
  • Amino Acid Sequence
  • Animals
  • Baculoviridae / genetics
  • Benzazepines / metabolism
  • Benzazepines / pharmacology
  • Cells, Cultured
  • Desensitization, Immunologic
  • Electrophoresis, Polyacrylamide Gel
  • Epitopes / genetics*
  • Fluorescent Antibody Technique
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Molecular Sequence Data
  • Moths
  • Palmitic Acid
  • Palmitic Acids / pharmacology
  • Phosphorylation
  • Proto-Oncogene Proteins c-myc / metabolism*
  • Receptors, Dopamine D1 / drug effects
  • Receptors, Dopamine D1 / genetics
  • Receptors, Dopamine D1 / metabolism*
  • Recombination, Genetic


  • Benzazepines
  • Epitopes
  • Palmitic Acids
  • Proto-Oncogene Proteins c-myc
  • Receptors, Dopamine D1
  • Palmitic Acid
  • 2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-benzazepine
  • Adenylyl Cyclases