Broad-scale phosphoprotein profiling of beta adrenergic receptor (β-AR) signaling reveals novel phosphorylation and dephosphorylation events

PloS One
Andrzej J ChruscinskiPaul J Utz

Abstract

β-adrenergic receptors (β-ARs) are model G-protein coupled receptors that mediate signal transduction in the sympathetic nervous system. Despite the widespread clinical use of agents that target β-ARs, the signaling pathways that operate downstream of β-AR stimulation have not yet been completely elucidated. Here, we utilized a lysate microarray approach to obtain a broad-scale perspective of phosphoprotein signaling downstream of β-AR. We monitored the time course of phosphorylation states of 54 proteins after β-AR activation mouse embryonic fibroblast (MEF) cells. In response to stimulation with the non-selective β-AR agonist isoproterenol, we observed previously described phosphorylation events such as ERK1/2(T202/Y204) and CREB(S133), but also novel phosphorylation events such as Cdc2(Y15) and Pyk2(Y402). All of these events were mediated through cAMP and PKA as they were reproduced by stimulation with the adenylyl cyclase activator forskolin and were blocked by treatment with H89, a PKA inhibitor. In addition, we also observed a number of novel isoproterenol-induced protein dephosphorylation events in target substrates of the PI3K/AKT pathway: GSK3β(S9), 4E-BP1(S65), and p70s6k(T389). These dephosphorylations were dependen...Continue Reading

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Citations

Aug 19, 2018·The Biochemical Journal·Blanca E Ruiz-MedinaRobert A Kirken
Jan 4, 2019·Cardiovascular Research·Nathalie MougenotLarissa Lipskaia
Jul 5, 2017·The Journal of Physiological Sciences : JPS·Ali KhalilimeybodiBabak Sharif-Kashani
May 15, 2018·Frontiers in Immunology·Thomas Sajda, Animesh A Sinha

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Methods Mentioned

BETA
Protein Assay
Pull-Down
Assay
Kinase
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Software Mentioned

SAM
Quantity One
GenePix Pro
GraphPad
TIGR Multiple Experiment Viewer ( TMEV )
Significance Analysis of Microarrays ( SAM )
GraphPad Prism

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