Specificity of receptor-G protein interactions. Discrimination of G(i) subtypes by the D2 dopamine receptor in a reconstituted system

S. E. Senogles, A. M. Spiegel, E. Padrell, R. Iyengar, M. G. Caron

Research output: Contribution to journalArticle

128 Scopus citations

Abstract

The selectivity of D2 dopamine receptor-guanine nucleotide-binding protein (G protein) coupling was studied by reconstitution techniques utilizing purified D2 dopamine receptors from bovine anterior pituitary and resolved G proteins from bovine brain, bovine pituitary, and human erythrocyte. Titration of a fixed receptor concentration with varying G protein concentrations revealed two aspects of receptor-G protein coupling. First, G(i2) appeared to couple selectively with the D2 receptor with ~ 10-fold higher affinity than any other tested G(i) subtype. Second, the G proteins differed in the maximal receptor-mediated agonist stimulation of the intrinsic GTPase activity. G(i2) appeared to be maximally stimulated by agonist-receptor complex with turnover numbers of ~ 2 min-1. The other G(i) subtypes, G(i1) and G(i3), could be only partially activated, resulting in maximal rates of GTPase of ~ 1 min-1. Agonist-stimulated GTPase activity was not detected in preparations containing G(o) from bovine brain. The differences in maximal agonist-stimulated GTPase rates observed among the G(i) subtypes could be explained by differences in agonist-promoted guanyl nucleotide exchange. Both guanosine 5'-3-O-(thio)triphosphate (GTPγS) binding and GDP release parameters were enhanced 2-fold for the G(i2) subtype over the other G(i) subtypes. These results suggest that even though several types of pertussis toxin substrate may exist in most tissues, a receptor may interact discretely with G proteins, thereby dictating signal transduction mechanisms.

Original languageEnglish (US)
Pages (from-to)4507-4514
Number of pages8
JournalJournal of Biological Chemistry
Volume265
Issue number8
StatePublished - Mar 30 1990
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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