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Switching of the coupling of the β2-adrenergic receptor to different G proteins by protein kinase A

Abstract

Many of the G-protein-coupled receptors for hormones that bind to the cell surface can signal to the interior of the cell through several different classes of G protein1,2,3,4. For example, although most of the actions of the prototype β2-adrenergic receptor are mediated through Gs proteins and the cyclic-AMP-dependent protein kinase (PKA) system5,6, β-adrenergic receptors can also couple to Gi proteins1,2. Here we investigate the mechanism that controls the specificity of this coupling. We show that in HEK293 cells, stimulation of mitogen-activated protein (MAP) kinase by the β2-adrenergic receptor is mediated by the βγ subunits of pertussis-toxin-sensitive G proteins through a pathway involving the non-receptor tyrosine kinase c-Src and the G protein Ras. Activation of this pathway by the β2-adrenergic receptor requires that the receptor be phosphorylated by PKA because it is blocked by H-89, an inhibitor of PKA. Additionally, a mutant of the receptor, which lacks the sites normally phosphorylated by PKA, can activate adenylyl cyclase5, the enzyme that generates cAMP, but not MAP kinase. Our results demonstrate that a mechanism previously shown to mediate uncoupling of the β2-adrenergic receptor from Gs and thus heterologous desensitization7 (PKA-mediated receptor phosphorylation), also serves to ‘switch’ coupling of this receptor from Gs to Gi and initiate a new set of signalling events.

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Figure 1: β-AR-mediated activation of MAP kinase.
Figure 2: Isoprenaline activation of MAPK is dependent on PTX-sensitive G proteins.
Figure 3: Effect of the cAMP-dependent protein kinase A inhibitor H-89 on MAP kinase activation.
Figure 4: PKA phosphorylation of the β-AR is required for isoprenaline-induced phosphorylation of MAPK.
Figure 5: Model for β2-AR-mediated G-protein switching to activate MAP kinase.

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Acknowledgements

We thank M. Holben and D. Addison for secretarial assistance.

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Correspondence to Robert J. Lefkowitz.

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Daaka, Y., Luttrell, L. & Lefkowitz, R. Switching of the coupling of the β2-adrenergic receptor to different G proteins by protein kinase A. Nature 390, 88–91 (1997). https://doi.org/10.1038/36362

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