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

Circ Res. 2016 Sep 30;119(8):931-43. doi: 10.1161/CIRCRESAHA.116.308964. Epub 2016 Aug 30.


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 because of an enhanced β2 adrenergic receptor 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 β2 adrenergic receptor signal by preventing PKA signal access to the myofilaments and to restore contractile response to adrenergic stimulation.

Conclusions: In hypertrophic rabbit myocytes, selectively enhanced β2 adrenergic receptor signaling toward the myofilaments contributes to elevated PKA activity and PKA phosphorylation of myofilament proteins. Reintroduction of caveolin-3 is able to confine β2 adrenergic receptor signaling and restore myocyte contractility in response to β adrenergic stimulation.

Keywords: adrenergic receptor; heart failure; myofibrils; phosphorylation; protein kinase A.

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Cells, Cultured
  • Cyclic AMP-Dependent Protein Kinases / metabolism*
  • Heart Failure / enzymology*
  • Heart Failure / genetics*
  • Myocytes, Cardiac / enzymology
  • Myofibrils / enzymology*
  • Myofibrils / genetics*
  • Phosphorylation / physiology
  • Rabbits


  • Cyclic AMP-Dependent Protein Kinases