Activation of the inhibitory GTP-binding protein of adenylate cyclase, G(i), by β-adrenergic receptors in reconstituted phospholipid vesicles

T. Asano, T. Katada, A. G. Gilman, E. M. Ross

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Abstract

β-Adrenergic receptors and the inhibitory GTP-binding protein, G(i) of the adenylate cyclase system were reconstituted into phospholipid vesicles by the method described previously for reconstituting receptors and the stimulatory GTP-binding protein G(s) (Brandt, D.R., Asano, T., Pedersen, S.E., and Ross, E.M. (1983) Biochemistry 22, 4357-4362). In the receptor-G(i) vesicles, β-adrenergic agonists stimulated both the high-affinity binding of guanosine 5'-O-(3-thiotriphosphate) (GTPγS) to G(i) and GTPase activity to an extent similar to that observed in vesicles containing β-adrenergic receptors and G(s). Stimulation required receptors and displayed appropriate β-adrenergic specificity. The prior treatment of receptor-G(i) vesicles with islet-activating protein (pertussis toxin) plus NAD markedly inhibited both the isoproterenol-stimulated binding of GTPγS and the isoproterenol-stimulated GTPase activity. No contamination of G(i) by G(s) was apparent. These data suggest that receptors that typically stimulate adenylate cyclase activity may also activate the inhibitory system, perhaps as one mechanism of desensitization.

Original languageEnglish (US)
Pages (from-to)9351-9354
Number of pages4
JournalJournal of Biological Chemistry
Volume259
Issue number15
StatePublished - 1984

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GTP Phosphohydrolases
Pertussis Toxin
Isoproterenol
GTP-Binding Proteins
Adenylyl Cyclases
Adrenergic Receptors
Phospholipids
Chemical activation
Guanosine 5'-O-(3-Thiotriphosphate)
Adrenergic Agonists
Biochemistry
NAD
Adrenergic Agents
Contamination

ASJC Scopus subject areas

  • Biochemistry

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Activation of the inhibitory GTP-binding protein of adenylate cyclase, G(i), by β-adrenergic receptors in reconstituted phospholipid vesicles. / Asano, T.; Katada, T.; Gilman, A. G.; Ross, E. M.

In: Journal of Biological Chemistry, Vol. 259, No. 15, 1984, p. 9351-9354.

Research output: Contribution to journalArticle

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AU - Ross, E. M.

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N2 - β-Adrenergic receptors and the inhibitory GTP-binding protein, G(i) of the adenylate cyclase system were reconstituted into phospholipid vesicles by the method described previously for reconstituting receptors and the stimulatory GTP-binding protein G(s) (Brandt, D.R., Asano, T., Pedersen, S.E., and Ross, E.M. (1983) Biochemistry 22, 4357-4362). In the receptor-G(i) vesicles, β-adrenergic agonists stimulated both the high-affinity binding of guanosine 5'-O-(3-thiotriphosphate) (GTPγS) to G(i) and GTPase activity to an extent similar to that observed in vesicles containing β-adrenergic receptors and G(s). Stimulation required receptors and displayed appropriate β-adrenergic specificity. The prior treatment of receptor-G(i) vesicles with islet-activating protein (pertussis toxin) plus NAD markedly inhibited both the isoproterenol-stimulated binding of GTPγS and the isoproterenol-stimulated GTPase activity. No contamination of G(i) by G(s) was apparent. These data suggest that receptors that typically stimulate adenylate cyclase activity may also activate the inhibitory system, perhaps as one mechanism of desensitization.

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