Systems analysis of PKA-mediated phosphorylation gradients in live cardiac myocytes

Jeffrey J. Saucerman, Jin Zhang, Jody C. Martin, Lili X. Peng, Antine E. Stenbit, Roger Y. Tsien, Andrew D. McCulloch

Research output: Contribution to journalArticle

Abstract

Compartmentation and dynamics of cAMP and PKA signaling are important determinants of specificity among cAMP's myriad cellular roles. Both cardiac inotropy and the progression of heart disease are affected by spatiotemporal variations in cAMP/PKA signaling, yet the dynamic patterns of PKA-mediated phosphorylation that influence differential responses to agonists have not been characterized. We performed live-cell imaging and systems modeling of PKA-mediated phosphorylation in neonatal cardiac myocytes in response to G-protein coupled receptor stimuli and UV photolysis of "caged" cAMP. cAMP accumulation was rate-limiting in PKA-mediated phosphorylation downstream of the β-adrenergic receptor. Prostaglandin E1 stimulated higher PKA activity in the cytosol than at the sarcolemma, whereas isoproterenol triggered faster sarcolemmal responses than cytosolic, likely due to restricted cAMP diffusion from submembrane compartments. Localized UV photolysis of caged cAMP triggered gradients of PKA-mediated phosphorylation, enhanced by phosphodiesterase activity and PKA-mediated buffering of cAMP. These findings indicate that combining live-cell FRET imaging and mechanistic computational models can provide quantitative understanding of spatiotemporal signaling.

Original languageEnglish (US)
Pages (from-to)12923-12928
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume103
Issue number34
DOIs
StatePublished - Aug 22 2006

Keywords

  • Fluorescence imaging
  • Models
  • Signal transduction

ASJC Scopus subject areas

  • Genetics
  • General

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    Saucerman, J. J., Zhang, J., Martin, J. C., Peng, L. X., Stenbit, A. E., Tsien, R. Y., & McCulloch, A. D. (2006). Systems analysis of PKA-mediated phosphorylation gradients in live cardiac myocytes. Proceedings of the National Academy of Sciences of the United States of America, 103(34), 12923-12928. https://doi.org/10.1073/pnas.0600137103