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J Biol Chem, Vol. 274, Issue 37, 26337-26343, September 10, 1999

Nitric Oxide Modulates ₂-Adrenergic Receptor Palmitoylation and Signaling

From the Département de Biochimie and Le Groupe de Recherche sur le Système Nerveux Autonome, Université de Montréal, Montréal, Quebec H3T 1J4, Canada and ^¶ Metabolic Diseases Branch, NIDDK, National Institutes of Health, Bethesda, Maryland 20892

To determine whether nitric oxide (NO) modulates the -adrenergic signaling pathway, we treated cells expressing ₂-adrenergic receptors (₂AR) with the NO donors, 3-morpholinosydnonimine (SIN-1) and 1,2,3,4-oxatriazolium,5-amino-3-(3-chloro-2-methylphenyl)chloride and determined the intracellular production of cAMP after exposure to -adrenergic receptor agonists, cholera toxin and forskolin. NO significantly decreased the potency of the -adrenergic agonist, isoproterenol, to stimulate cAMP production without affecting the stimulatory action of forskolin and cholera toxin, which directly activate adenylyl cyclase and G_s, respectively. Treatment with the NO donor increased the guanyl nucleotide-sensitive high affinity constant for the agonist, isoproterenol, thus suggesting that it reduced functional coupling between the receptor and G_s. Stimulation of endogenous NO production by lipopolysaccharide in RAW 264.7 macrophages also caused a significant increase in the EC₅₀ for isoproterenol-stimulated cAMP production. SIN-1 treatment also led to a reduction in both basal and isoproterenol-stimulated incorporation of [³H]palmitate into the ₂AR. Signaling through the nonpalmitoylated, Gly³⁴¹₂AR mutant was unchanged by SIN-1 treatment. Given the link between ₂AR palmitoylation and its responsiveness to agonist, these results suggest that the primary action of NO was depalmitoylation of the ₂AR resulting in decreased signaling through the ₂AR.

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