Specificity of receptor-G protein interactions

Discrimination of Gi subtypes by the D2 dopamine receptor in a reconstituted system

Susan E. Senogles, Allen M. Spiegel, Elena Padrell, Ravi Iyengar, Marc G. Caron

Research output: Contribution to journalArticle

126 Citations (Scopus)

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, Gi2 appeared to couple selectively with the D2 receptor with ∼10-fold higher affinity than any other tested Gi subtype. Second, the G proteins differed in the maximal receptor-mediated agonist stimulation of the intrinsic GTPase activity. Gi2 appeared to be maximally stimulated by agonist-receptor complex with turnover numbers of ∼2 min-1. The other Gi subtypes, Gi1 and Gi3, 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 Go from bovine brain. The differences in maximal agonist-stimulated GTPase rates observed among the Gi 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 Gi2 subtype over the other Gi 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 15 1990
Externally publishedYes

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Dopamine D2 Receptors
GTP-Binding Proteins
GTP Phosphohydrolases
Brain
Guanosine 5'-O-(3-Thiotriphosphate)
Signal transduction
Guanine Nucleotides
Pertussis Toxin
Titration
Signal Transduction
Carrier Proteins
Nucleotides
Erythrocytes
Tissue
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Specificity of receptor-G protein interactions : Discrimination of Gi subtypes by the D2 dopamine receptor in a reconstituted system. / Senogles, Susan E.; Spiegel, Allen M.; Padrell, Elena; Iyengar, Ravi; Caron, Marc G.

In: Journal of Biological Chemistry, Vol. 265, No. 8, 15.03.1990, p. 4507-4514.

Research output: Contribution to journalArticle

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