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

Overview

Journal Circ Res

Specialty Cardiology & Vascular Diseases

Date 2016 Sep 1

PMID 27576469

Citations 35

Authors

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 Bossuyt

Donald Bers

Yang K Xiang

Affiliations

Soon will be listed here.

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 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.

Citing Articles

Caveolin and oxidative stress in cardiac pathology.

Zadorozny L, Du J, Supanekar N, Annamalai K, Yu Q, Wang M Front Physiol. 2025; 16:1550647.

PMID: 40041164 PMC: 11876135. DOI: 10.3389/fphys.2025.1550647.

Carvedilol Activates a Myofilament Signaling Circuitry to Restore Cardiac Contractility in Heart Failure.

Wang Y, Zhao M, Liu X, Xu B, Reddy G, Jovanovic A JACC Basic Transl Sci. 2024; 9(8):982-1001.

PMID: 39297139 PMC: 11405995. DOI: 10.1016/j.jacbts.2024.03.007.

Differential Downregulation of β-Adrenergic Receptor Signaling in the Heart.

Xu B, Bahriz S, Salemme V, Wang Y, Zhu C, Zhao M J Am Heart Assoc. 2024; 13(12):e033733.

PMID: 38860414 PMC: 11255761. DOI: 10.1161/JAHA.123.033733.

Abnormal phosphorylation / dephosphorylation and Ca dysfunction in heart failure.

Liu Y, Wang Q, Song Y, Song X, Fan Y, Kong L Heart Fail Rev. 2024; 29(4):751-768.

PMID: 38498262 DOI: 10.1007/s10741-024-10395-w.

Phosphodiesterase in heart and vessels: from physiology to diseases.

Fu Q, Wang Y, Yan C, Xiang Y Physiol Rev. 2023; 104(2):765-834.

PMID: 37971403 PMC: 11281825. DOI: 10.1152/physrev.00015.2023.

References

Bryant S, Kimura T, Kong C, Watson J, Chase A, Suleiman M . Stimulation of ICa by basal PKA activity is facilitated by caveolin-3 in cardiac ventricular myocytes. J Mol Cell Cardiol. 2014; 68:47-55. PMC: 3980375. DOI: 10.1016/j.yjmcc.2013.12.026. View

Calaghan S, Kozera L, White E . Compartmentalisation of cAMP-dependent signalling by caveolae in the adult cardiac myocyte. J Mol Cell Cardiol. 2008; 45(1):88-92. DOI: 10.1016/j.yjmcc.2008.04.004. View

Wright P, Nikolaev V, OHara T, Diakonov I, Bhargava A, Tokar S . Caveolin-3 regulates compartmentation of cardiomyocyte beta2-adrenergic receptor-mediated cAMP signaling. J Mol Cell Cardiol. 2013; 67:38-48. PMC: 4266930. DOI: 10.1016/j.yjmcc.2013.12.003. View

Messer A, Marston S . Investigating the role of uncoupling of troponin I phosphorylation from changes in myofibrillar Ca(2+)-sensitivity in the pathogenesis of cardiomyopathy. Front Physiol. 2014; 5:315. PMC: 4142463. DOI: 10.3389/fphys.2014.00315. View

Gresham K, Stelzer J . The contributions of cardiac myosin binding protein C and troponin I phosphorylation to β-adrenergic enhancement of in vivo cardiac function. J Physiol. 2015; 594(3):669-86. PMC: 4930068. DOI: 10.1113/JP270959. View

Markandeya Y, Phelan L, Woon M, Keefe A, Reynolds C, August B . Caveolin-3 Overexpression Attenuates Cardiac Hypertrophy via Inhibition of T-type Ca2+ Current Modulated by Protein Kinase Cα in Cardiomyocytes. J Biol Chem. 2015; 290(36):22085-100. PMC: 4571960. DOI: 10.1074/jbc.M115.674945. View

Ruehr M, Russell M, Bond M . A-kinase anchoring protein targeting of protein kinase A in the heart. J Mol Cell Cardiol. 2004; 37(3):653-65. DOI: 10.1016/j.yjmcc.2004.04.017. View

Fraysse B, Weinberger F, Bardswell S, Cuello F, Vignier N, Geertz B . Increased myofilament Ca2+ sensitivity and diastolic dysfunction as early consequences of Mybpc3 mutation in heterozygous knock-in mice. J Mol Cell Cardiol. 2012; 52(6):1299-307. PMC: 3370652. DOI: 10.1016/j.yjmcc.2012.03.009. View

Perrino C, Naga Prasad S, Schroder J, Hata J, Milano C, Rockman H . Restoration of beta-adrenergic receptor signaling and contractile function in heart failure by disruption of the betaARK1/phosphoinositide 3-kinase complex. Circulation. 2005; 111(20):2579-87. DOI: 10.1161/CIRCULATIONAHA.104.508796. View

10.

Fu Q, Chen X, Xiang Y . Compartmentalization of β-adrenergic signals in cardiomyocytes. Trends Cardiovasc Med. 2013; 23(7):250-6. PMC: 4264830. DOI: 10.1016/j.tcm.2013.02.001. View

11.

Rockman H, Koch W, Lefkowitz R . Seven-transmembrane-spanning receptors and heart function. Nature. 2002; 415(6868):206-12. DOI: 10.1038/415206a. View

12.

Dong X, Sumandea C, Chen Y, Garcia-Cazarin M, Zhang J, Balke C . Augmented phosphorylation of cardiac troponin I in hypertensive heart failure. J Biol Chem. 2011; 287(2):848-57. PMC: 3256890. DOI: 10.1074/jbc.M111.293258. View

13.

Mehel H, Emons J, Vettel C, Wittkopper K, Seppelt D, Dewenter M . Phosphodiesterase-2 is up-regulated in human failing hearts and blunts β-adrenergic responses in cardiomyocytes. J Am Coll Cardiol. 2013; 62(17):1596-606. DOI: 10.1016/j.jacc.2013.05.057. View

14.

De Arcangelis V, Liu S, Zhang D, Soto D, Xiang Y . Equilibrium between adenylyl cyclase and phosphodiesterase patterns adrenergic agonist dose-dependent spatiotemporal cAMP/protein kinase A activities in cardiomyocytes. Mol Pharmacol. 2010; 78(3):340-9. PMC: 2939479. DOI: 10.1124/mol.110.064444. View

15.

Freeman K, Olsson M, Moore R, Weinberger H, GRUPP I, Vikstrom K . Progression from hypertrophic to dilated cardiomyopathy in mice that express a mutant myosin transgene. Am J Physiol Heart Circ Physiol. 2000; 280(1):H151-9. DOI: 10.1152/ajpheart.2001.280.1.H151. View

16.

Balijepalli R, Foell J, Hall D, Hell J, Kamp T . Localization of cardiac L-type Ca(2+) channels to a caveolar macromolecular signaling complex is required for beta(2)-adrenergic regulation. Proc Natl Acad Sci U S A. 2006; 103(19):7500-5. PMC: 1564282. DOI: 10.1073/pnas.0503465103. View

17.

Galbiati F, Volonte D, CHU J, Li M, Fine S, Fu M . Transgenic overexpression of caveolin-3 in skeletal muscle fibers induces a Duchenne-like muscular dystrophy phenotype. Proc Natl Acad Sci U S A. 2000; 97(17):9689-94. PMC: 16926. DOI: 10.1073/pnas.160249097. View

18.

Pogwizd S, Qi M, Yuan W, Samarel A, Bers D . Upregulation of Na(+)/Ca(2+) exchanger expression and function in an arrhythmogenic rabbit model of heart failure. Circ Res. 1999; 85(11):1009-19. DOI: 10.1161/01.res.85.11.1009. View

19.

van der Velden J, Narolska N, Lamberts R, Boontje N, Borbely A, Zaremba R . Functional effects of protein kinase C-mediated myofilament phosphorylation in human myocardium. Cardiovasc Res. 2005; 69(4):876-87. DOI: 10.1016/j.cardiores.2005.11.021. View

20.

Koss K, Kranias E . Phospholamban: a prominent regulator of myocardial contractility. Circ Res. 1996; 79(6):1059-63. DOI: 10.1161/01.res.79.6.1059. View