Genetically Encoded Biosensors Reveal PKA Hyperphosphorylation on the Myofilaments in Rabbit Heart Failure

Federica Barbagallo, Bing Xu, Gopireddy R. Reddy, Toni West, Qingtong Wang, Qin Fu, Minghui Li, Qian Shi, Kenneth S Ginsburg, William Ferrier, Andrea M. Isidori, Fabio Naro, Hemal H. Patel, Julie B C Bossuyt, Donald M Bers, Yang Kevin Xiang

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

RATIONALE:: In heart failure, myofilament proteins display abnormal phosphorylation, which contributes to contractile dysfunction. The mechanisms underlying the dysregulation of protein phosphorylation on myofilaments is not clear. OBJECTIVE:: This study aims to understand the mechanisms underlying altered phosphorylation of myofilament proteins in heart failure. METHODS AND RESULTS:: We generate a novel genetically encoded protein kinase A (PKA) biosensor anchored onto the myofilaments in rabbit cardiac myocytes to examine PKA activity at the myofilaments in responses to adrenergic stimulation. We show that PKA activity is shifted from the sarcolemma to the myofilaments in hypertrophic failing rabbit myocytes. In particular, the increased PKA activity on the myofilaments is due to an enhanced β2 adrenergic receptor (β2AR) signal selectively directed to the myofilaments together with a reduced phosphodiesterase activity associated with the myofibrils. Mechanistically, the enhanced PKA activity on the myofilaments is associated with downregulation of caveolin-3 in the hypertrophic failing rabbit myocytes. Reintroduction of caveolin-3 in the failing myocytes is able to normalize the distribution of β2AR signal by preventing PKA signal access to the myofilaments, and to restore contractile response to adrenergic stimulation. CONCLUSIONS:: In hypertrophic rabbit myocytes, selectively enhanced β2AR signaling toward the myofilaments contributes to elevated PKA activity and PKA phosphorylation of myofilament proteins. Reintroduction of caveolin-3 is able to confine β2AR signaling and restore myocyte contractility in response to β-adrenergic stimulation.

Original languageEnglish (US)
JournalCirculation Research
DOIs
StateAccepted/In press - Aug 31 2016

ASJC Scopus subject areas

  • Physiology
  • Medicine(all)
  • Cardiology and Cardiovascular Medicine

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    Barbagallo, F., Xu, B., Reddy, G. R., West, T., Wang, Q., Fu, Q., Li, M., Shi, Q., Ginsburg, K. S., Ferrier, W., Isidori, A. M., Naro, F., Patel, H. H., Bossuyt, J. B. C., Bers, D. M., & Xiang, Y. K. (Accepted/In press). Genetically Encoded Biosensors Reveal PKA Hyperphosphorylation on the Myofilaments in Rabbit Heart Failure. Circulation Research. https://doi.org/10.1161/CIRCRESAHA.116.308964