Reconstitution of catecholamine-stimulated adenylate cyclase activity using three purified proteins

D. C. May, E. M. Ross, A. G. Gilman, M. D. Smigel

Research output: Contribution to journalArticle

103 Citations (Scopus)

Abstract

β-Adrenergic receptors, the GTP-binding regulatory protein that stimulates adenylate cyclase (G(s)), and adenylate cyclase were each purified and reconstituted into unilamellar vesicles composed of phosphatidylethanolamine and phosphatidylserine (3:2, w/w). The molar ratio of receptor:G(s):adenylate cyclase was estimated to be about 1:10:1. Adenylate cyclase activity in the vesicles was stimulated up to 2.6-fold by β-adrenergic agonists. Stimulation was dependent on the presence of guanine nucleotide, displayed appropriate β-adrenergic selectivity and stereoselectivity for agonists, and was blocked appropriately by β-adrenergic antagonists. Therefore, while additional proteins may modulate adenylate cyclase activity in native membranes, these results show that these three proteins are sufficient for the expression of hormone-stimulated adenylate cyclase.

Original languageEnglish (US)
Pages (from-to)15829-15833
Number of pages5
JournalJournal of Biological Chemistry
Volume260
Issue number29
StatePublished - 1985

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Adenylyl Cyclases
Catecholamines
Proteins
GTP-Binding Proteins
Stereoselectivity
Unilamellar Liposomes
Adrenergic Agonists
Adrenergic Antagonists
Guanine Nucleotides
Phosphatidylserines
Guanosine Triphosphate
Adrenergic Agents
Adrenergic Receptors
Hormones
Membranes

ASJC Scopus subject areas

  • Biochemistry

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Reconstitution of catecholamine-stimulated adenylate cyclase activity using three purified proteins. / May, D. C.; Ross, E. M.; Gilman, A. G.; Smigel, M. D.

In: Journal of Biological Chemistry, Vol. 260, No. 29, 1985, p. 15829-15833.

Research output: Contribution to journalArticle

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