Expression and characterization of the rat D3 dopamine receptor: Pharmacologic properties and development of antibodies

V. A. Boundy, R. R. Luedtke, A. L. Gallitano, J. E. Smith, T. M. Filtz, R. G. Kallen, P. B. Molinoff

Research output: Contribution to journalArticle

83 Scopus citations

Abstract

A baculovirus expression system provided an enriched source of biologically and immunologically active D3 dopamine receptors. Receptors expressed in Spodoptera frugiperda insect (Sf9) cells at a density of 5 to 15 pmol/mg of protein displayed high affinity for the antagonists, eticlopride, fluphenazine and spiroperidol, and the agonist, N-propylnorapomorphine. The binding of agonists was not sensitive to GTP. Antisera raised against synthetic peptides in the third intracellular loop of the D3 dopamine receptor immunoprecipitated binding sites for (S)-3-[125I]-iodo-2- hydroxy-5,6-dimethoxy-N-[(1-ethyl-2-pyrrolidinyl)-methyl]-benz amide from solubilized extracts of infected Sf9 cells and detergent extracts of rat caudate. These antisera specifically recognized a single band on immunoblots of Sf9 cells infected with recombinant D3 baculovirus. Both the immunoprecipitation and immunoblot reactions were blocked by preincubation of the antisera with the immunization peptide. These results suggest that the D3 receptor protein is expressed in rat brain.

Original languageEnglish (US)
Pages (from-to)1002-1011
Number of pages10
JournalJournal of Pharmacology and Experimental Therapeutics
Volume264
Issue number2
StatePublished - Dec 1 1993
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology

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