Increased palmitoylation of the Gs protein α subunit after activation by the β-adrenergic receptor or cholera toxin

Michael Y. Degtyarev, Allen M. Spiegel, Teresa L Z Jones

Research output: Contribution to journalArticle

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Abstract

The α subunit of the heterotrimeric Gs protein that couples the β-adrenergic receptor to adenylyl cyclase undergoes post-translational palmitoylation. We examined the dynamics of this modification of αs by metabolic labeling of COS and S49 lymphoma cells under different conditions. The endogenous αs proteins were immunoprecipitated with a peptide-specific antibody, separated by SDS-polyacrylamide gel electrophoresis, and analyzed by fluorography and densitometry. A pulse-chase study of COS cells incubated with [3H]palmitate or [35S]methionine showed that for αs the palmitate turnover (t1/2 ≈ 50 min) was significantly faster than the protein degradation. Treatment of cells with 10 μM isoproterenol, a β-adrenergic receptor agonist, in the presence of [3H]palmitate led to a rapid 4-10-fold increase in the palmitoylation of αs. This increase in palmitoylation was concentration-dependent (EC50 = 0.9 μM) and blocked by the antagonist propranolol. The mutant αs proteins in the unc and H21a S49 cell lines did not show an increase in [3H]palmitate incorporation with isoproterenol treatment. Cholera toxin treatment of COS cells increased the [3H]palmitate incorporation into the αs subunits. These data indicate that palmitoylation of the αs subunit is dynamic and regulated by activation of the αs subunit.

Original languageEnglish (US)
Pages (from-to)23769-23772
Number of pages4
JournalJournal of Biological Chemistry
Volume268
Issue number32
StatePublished - Nov 15 1993
Externally publishedYes

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Lipoylation
Palmitates
Cholera Toxin
Protein Subunits
Adrenergic Receptors
Chemical activation
COS Cells
Isoproterenol
Photofluorography
Proteins
Cells
Adrenergic Agonists
Densitometry
Mutant Proteins
Electrophoresis
Adenylyl Cyclases
Propranolol
Methionine
Labeling
Proteolysis

ASJC Scopus subject areas

  • Biochemistry

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Increased palmitoylation of the Gs protein α subunit after activation by the β-adrenergic receptor or cholera toxin. / Degtyarev, Michael Y.; Spiegel, Allen M.; Jones, Teresa L Z.

In: Journal of Biological Chemistry, Vol. 268, No. 32, 15.11.1993, p. 23769-23772.

Research output: Contribution to journalArticle

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