Construction of a soluble adenylyl cyclase activated by Gsα and forskolin

Wei Jen Tang, Alfred G. Gilman

Research output: Contribution to journalArticle

159 Citations (Scopus)

Abstract

A soluble adenylyl cyclase was constructed by linkage of portions of the cytosolic domains of the mammalian type I and type II enzymes. The soluble enzyme was stimulated by both forskolin and the α subunit of the heterotrimeric guanine nucleotidebinding protein (G protein) Gs (Gsα). Expression of the construct complemented the catabolic defect in a strain of Escherichia coli that is deficient in adenylyl cyclase activity. The active, approximately 60-kilodalton enzyme accumulated in the cytoplasmic fraction of E. coli to yield activities in excess of 1 nanomole per minute per milligram of protein. The two sets of transmembrane helices of mammalian adenylyl cyclases are thus not necessary for the catalytic or the most characteristic regulatory activities of the enzyme. This system may be useful for both genetic and biochemical analysis of G protein-regulated adenylyl cyclases.

Original languageEnglish (US)
Pages (from-to)1769-1772
Number of pages4
JournalScience
Volume268
Issue number5218
StatePublished - Jun 23 1995

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Colforsin
Adenylyl Cyclases
Enzymes
GTP-Binding Proteins
Escherichia coli
Guanine
Molecular Biology
Proteins

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  • General

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Tang, W. J., & Gilman, A. G. (1995). Construction of a soluble adenylyl cyclase activated by Gsα and forskolin. Science, 268(5218), 1769-1772.

Construction of a soluble adenylyl cyclase activated by Gsα and forskolin. / Tang, Wei Jen; Gilman, Alfred G.

In: Science, Vol. 268, No. 5218, 23.06.1995, p. 1769-1772.

Research output: Contribution to journalArticle

Tang, WJ & Gilman, AG 1995, 'Construction of a soluble adenylyl cyclase activated by Gsα and forskolin', Science, vol. 268, no. 5218, pp. 1769-1772.
Tang WJ, Gilman AG. Construction of a soluble adenylyl cyclase activated by Gsα and forskolin. Science. 1995 Jun 23;268(5218):1769-1772.
Tang, Wei Jen ; Gilman, Alfred G. / Construction of a soluble adenylyl cyclase activated by Gsα and forskolin. In: Science. 1995 ; Vol. 268, No. 5218. pp. 1769-1772.
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