Expression and characterization of the rat D3 dopamine receptor

Pharmacologic properties and development of antibodies

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

Research output: Contribution to journalArticle

83 Citations (Scopus)

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 - 1993
Externally publishedYes

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Dopamine D3 Receptors
Sf9 Cells
Immune Sera
eticlopride
Baculoviridae
Antibodies
Fluphenazine
Spiperone
Spodoptera
Peptides
Guanosine Triphosphate
Cell Extracts
Immunoprecipitation
Amides
Detergents
Insects
Immunization
Proteins
Binding Sites
Brain

ASJC Scopus subject areas

  • Pharmacology

Cite this

Expression and characterization of the rat D3 dopamine receptor : Pharmacologic properties and development of antibodies. / Boundy, V. A.; Luedtke, R. R.; Gallitano-Mendel, Amelia L; Smith, J. E.; Filtz, T. M.; Kallen, R. G.; Molinoff, P. B.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 264, No. 2, 1993, p. 1002-1011.

Research output: Contribution to journalArticle

Boundy, V. A. ; Luedtke, R. R. ; Gallitano-Mendel, Amelia L ; Smith, J. E. ; Filtz, T. M. ; Kallen, R. G. ; Molinoff, P. B. / Expression and characterization of the rat D3 dopamine receptor : Pharmacologic properties and development of antibodies. In: Journal of Pharmacology and Experimental Therapeutics. 1993 ; Vol. 264, No. 2. pp. 1002-1011.
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