Abstract
Activation of the β adrenergic receptor (βAR) induces a tightly controlled cAMP/protein kinase A (PKA) activity to ensure an agonist dose-dependent and saturable contraction response in animal heart. We have found that stimulation of β1AR by isoproterenol induces maximal contraction responses at the dose of 1 μM in cardiac myocytes; however, cAMP accumulation continues to increase with higher agonist concentrations. Dose-dependent cAMP accumulation is tightly controlled by negative regulator phosphodiesterase 4 (PDE4) that hydrolyzes cAMP. At 1 nM isoproterenol, cAMP accumulation is minimal because of the hydrolysis of cAMP by PDE4, which leads to a small increase in PKA phosphorylation of phospholamban and troponin I (TnI), and contraction responses. Inhibition of PDE4 activity with rolipram enhances cAMP accumulation, yields maximal PKA phosphorylation of phospholamban and TnI, and myocyte contraction responses. In contrast, at 10 μM isoproterenol, despite the negative effect of PDE4, cAMP accumulation is sufficient for maximal PKA phosphorylation of phospholamban and TnI. Inhibition of PDE4 with rolipram enhances cAMP accumulation, but not PKA phosphorylation and contraction responses. It is interesting that activities of both PKA and protein phosphatase 2A (PP2A) are enhanced under β1AR activation with 10 μM isoproterenol, and PP2A is recruited to PKA/A kinase-anchoring protein complex. Inhibition of PP2A with okadaic acid further enhances the phosphorylation of phospholamban and TnI as well as contraction responses induced by 10 μM isoproterenol. Therefore, PP2A plays a key role in limiting PKA phosphorylation of phospholamban and TnI for myocyte contraction responses under β1AR stimulation.
Footnotes
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This work was supported by grants from the National Institutes of Health and the American Heart Association (to Y.X.).
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ABBREVIATIONS: βAR, β-adrenergic receptor; TnI, troponin I, PDE, phosphodiesterase; AKAP, A kinase anchoring proteins; IBMX, 3-isoproterenolbutyl-1-methylxanthine; PKA, protein kinase A; PKI, protein kinase A inhibitor; KO, knockout; PP2A, protein phosphatase 2A.
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The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.
- Received June 17, 2008.
- Accepted August 13, 2008.
- The American Society for Pharmacology and Experimental Therapeutics
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