Biochemical characterization of three stimulatory GTP-binding proteins: The large and small forms of Gs and the olfactory-specific G-protein, Golf

David T. Jones, Susan B. Masters, Henry R. Bourne, Randall R Reed

Research output: Contribution to journalArticle

Abstract

The biochemical properties of three stimulatory guanine nucleotide-binding protein (G-protein) a subunits, the large and small forms of Gs, Gs-l (52 kDa) and Gs-8 (45 kDa), and the olfactory specific G-protein, Golf, have been compared. Complementary DNAs (cDNAs) encoding each α subunit were independently expressed in a mammalian cell line deficient in endogenous stimulatory G-proteins (S49 cyc-kin-). Gs-l and Gs-8 respond similarly to activation by the β-adrenergic agonist isoproterenol (EC50 = 80 and 60 nM, respectively) and the receptor-independent G-protein activators guanosine 5-O-3-(thio)triphosphate) (GTPγS) and AlF4-. The ability of Golf to interact with the β-adrenergic receptor was also examined. Surprisingly, Golf interacts with β-adrenergic receptors and is activated by isoproterenol (EC50 = 120 nM). All three G-proteins respond similarly to treatment with different α, β, and γ thiophosphoryl analogs of GTP. Specifically, (R)-GTPαS and GTPγS activate each G-protein, whereas (S)-GTPαS and (R)-or (S)-GTPβS are inactive. In addition, similar to G, Golfα is covalently modified and constitutively activated by cholera toxin. These studies demonstrate that all three stimulatory G-proteins are functionally and structurally similar, however, subtle differences between Golf and the two forms of Gs appear to modulate their interactions with receptors.

Original languageEnglish (US)
Pages (from-to)2671-2676
Number of pages6
JournalJournal of Biological Chemistry
Volume265
Issue number5
StatePublished - 1990

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Golf
Guanine Nucleotides
GTP-Binding Proteins
Carrier Proteins
Isoproterenol
Adrenergic Receptors
Guanosine 5'-O-(3-Thiotriphosphate)
Adrenergic Agonists
Cholera Toxin
Protein S
Guanosine Triphosphate
Protein Subunits
Complementary DNA
Chemical activation
Cells
Cell Line

ASJC Scopus subject areas

  • Biochemistry

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Biochemical characterization of three stimulatory GTP-binding proteins : The large and small forms of Gs and the olfactory-specific G-protein, Golf. / Jones, David T.; Masters, Susan B.; Bourne, Henry R.; Reed, Randall R.

In: Journal of Biological Chemistry, Vol. 265, No. 5, 1990, p. 2671-2676.

Research output: Contribution to journalArticle

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