HNO Enhances SERCA2a activity and cardiomyocyte function by promoting redox-dependent phospholamban oligomerization

Vidhya Sivakumaran, Brian A. Stanley, Carlo G. Tocchetti, Jeff D. Ballin, Viviane Caceres, Lufang Zhou, Gizem Keceli, Peter P. Rainer, Dong Lee, Sabine Huke, Mark T. Ziolo, Evangelia G. Kranias, John P. Toscano, Gerald M. Wilson, Brian O'Rourke, David A Kass, James E. Mahaney, Nazareno Paolocci

Research output: Contribution to journalArticle

Abstract

Aims: Nitroxyl (HNO) interacts with thiols to act as a redox-sensitive modulator of protein function. It enhances sarcoplasmic reticular Ca 2+ uptake and myofilament Ca2+ sensitivity, improving cardiac contractility. This activity has led to clinical testing of HNO donors for heart failure. Here we tested whether HNO alters the inhibitory interaction between phospholamban (PLN) and the sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) in a redox-dependent manner, improving Ca 2+ handling in isolated myocytes/hearts. Results: Ventriculocytes, sarcoplasmic reticulum (SR) vesicles, and whole hearts were isolated from control (wildtype [WT]) or PLN knockout (pln-/-) mice. Compared to WT, pln-/- myocytes displayed enhanced resting sarcomere shortening, peak Ca2+ transient, and blunted β-adrenergic responsiveness. HNO stimulated shortening, relaxation, and Ca2+ transient in WT cardiomyocytes, and evoked positive inotropy/lusitropy in intact hearts. These changes were markedly blunted in pln-/- cells/hearts. HNO enhanced SR Ca2+ uptake in WT but not pln-/- SR-vesicles. Spectroscopic studies in insect cell microsomes expressing SERCA2a±PLN showed that HNO increased Ca2+-dependent SERCA2a conformational flexibility but only when PLN was present. In cardiomyocytes, HNO achieved this effect by stabilizing PLN in an oligomeric disulfide bond-dependent configuration, decreasing the amount of free inhibitory monomeric PLN available. Innovation: HNO-dependent redox changes in myocyte PLN oligomerization relieve PLN inhibition of SERCA2a. Conclusions: PLN plays a central role in HNO-induced enhancement of SERCA2a activity, leading to increased inotropy/lusitropy in intact myocytes and hearts. PLN remains physically associated with SERCA2a; however, less monomeric PLN is available resulting in decreased inhibition of the enzyme. These findings offer new avenues to improve Ca2+ handling in failing hearts. Antioxid. Redox Signal. 19, 1185-1197.

Original languageEnglish (US)
Pages (from-to)1185-1197
Number of pages13
JournalAntioxidants and Redox Signaling
Volume19
Issue number11
DOIs
StatePublished - Oct 10 2013

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Oligomerization
Cardiac Myocytes
Oxidation-Reduction
Muscle Cells
Sarcoplasmic Reticulum
Sarcoplasmic Reticulum Calcium-Transporting ATPases
phospholamban
Sarcomeres
Myofibrils
Microsomes
Sulfhydryl Compounds
Knockout Mice
Disulfides
Adrenergic Agents
Modulators
Insects
Heart Failure
Innovation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology
  • Physiology
  • Clinical Biochemistry

Cite this

HNO Enhances SERCA2a activity and cardiomyocyte function by promoting redox-dependent phospholamban oligomerization. / Sivakumaran, Vidhya; Stanley, Brian A.; Tocchetti, Carlo G.; Ballin, Jeff D.; Caceres, Viviane; Zhou, Lufang; Keceli, Gizem; Rainer, Peter P.; Lee, Dong; Huke, Sabine; Ziolo, Mark T.; Kranias, Evangelia G.; Toscano, John P.; Wilson, Gerald M.; O'Rourke, Brian; Kass, David A; Mahaney, James E.; Paolocci, Nazareno.

In: Antioxidants and Redox Signaling, Vol. 19, No. 11, 10.10.2013, p. 1185-1197.

Research output: Contribution to journalArticle

Sivakumaran, V, Stanley, BA, Tocchetti, CG, Ballin, JD, Caceres, V, Zhou, L, Keceli, G, Rainer, PP, Lee, D, Huke, S, Ziolo, MT, Kranias, EG, Toscano, JP, Wilson, GM, O'Rourke, B, Kass, DA, Mahaney, JE & Paolocci, N 2013, 'HNO Enhances SERCA2a activity and cardiomyocyte function by promoting redox-dependent phospholamban oligomerization', Antioxidants and Redox Signaling, vol. 19, no. 11, pp. 1185-1197. https://doi.org/10.1089/ars.2012.5057
Sivakumaran, Vidhya ; Stanley, Brian A. ; Tocchetti, Carlo G. ; Ballin, Jeff D. ; Caceres, Viviane ; Zhou, Lufang ; Keceli, Gizem ; Rainer, Peter P. ; Lee, Dong ; Huke, Sabine ; Ziolo, Mark T. ; Kranias, Evangelia G. ; Toscano, John P. ; Wilson, Gerald M. ; O'Rourke, Brian ; Kass, David A ; Mahaney, James E. ; Paolocci, Nazareno. / HNO Enhances SERCA2a activity and cardiomyocyte function by promoting redox-dependent phospholamban oligomerization. In: Antioxidants and Redox Signaling. 2013 ; Vol. 19, No. 11. pp. 1185-1197.
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abstract = "Aims: Nitroxyl (HNO) interacts with thiols to act as a redox-sensitive modulator of protein function. It enhances sarcoplasmic reticular Ca 2+ uptake and myofilament Ca2+ sensitivity, improving cardiac contractility. This activity has led to clinical testing of HNO donors for heart failure. Here we tested whether HNO alters the inhibitory interaction between phospholamban (PLN) and the sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) in a redox-dependent manner, improving Ca 2+ handling in isolated myocytes/hearts. Results: Ventriculocytes, sarcoplasmic reticulum (SR) vesicles, and whole hearts were isolated from control (wildtype [WT]) or PLN knockout (pln-/-) mice. Compared to WT, pln-/- myocytes displayed enhanced resting sarcomere shortening, peak Ca2+ transient, and blunted β-adrenergic responsiveness. HNO stimulated shortening, relaxation, and Ca2+ transient in WT cardiomyocytes, and evoked positive inotropy/lusitropy in intact hearts. These changes were markedly blunted in pln-/- cells/hearts. HNO enhanced SR Ca2+ uptake in WT but not pln-/- SR-vesicles. Spectroscopic studies in insect cell microsomes expressing SERCA2a±PLN showed that HNO increased Ca2+-dependent SERCA2a conformational flexibility but only when PLN was present. In cardiomyocytes, HNO achieved this effect by stabilizing PLN in an oligomeric disulfide bond-dependent configuration, decreasing the amount of free inhibitory monomeric PLN available. Innovation: HNO-dependent redox changes in myocyte PLN oligomerization relieve PLN inhibition of SERCA2a. Conclusions: PLN plays a central role in HNO-induced enhancement of SERCA2a activity, leading to increased inotropy/lusitropy in intact myocytes and hearts. PLN remains physically associated with SERCA2a; however, less monomeric PLN is available resulting in decreased inhibition of the enzyme. These findings offer new avenues to improve Ca2+ handling in failing hearts. Antioxid. Redox Signal. 19, 1185-1197.",
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AU - Sivakumaran, Vidhya

AU - Stanley, Brian A.

AU - Tocchetti, Carlo G.

AU - Ballin, Jeff D.

AU - Caceres, Viviane

AU - Zhou, Lufang

AU - Keceli, Gizem

AU - Rainer, Peter P.

AU - Lee, Dong

AU - Huke, Sabine

AU - Ziolo, Mark T.

AU - Kranias, Evangelia G.

AU - Toscano, John P.

AU - Wilson, Gerald M.

AU - O'Rourke, Brian

AU - Kass, David A

AU - Mahaney, James E.

AU - Paolocci, Nazareno

PY - 2013/10/10

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N2 - Aims: Nitroxyl (HNO) interacts with thiols to act as a redox-sensitive modulator of protein function. It enhances sarcoplasmic reticular Ca 2+ uptake and myofilament Ca2+ sensitivity, improving cardiac contractility. This activity has led to clinical testing of HNO donors for heart failure. Here we tested whether HNO alters the inhibitory interaction between phospholamban (PLN) and the sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) in a redox-dependent manner, improving Ca 2+ handling in isolated myocytes/hearts. Results: Ventriculocytes, sarcoplasmic reticulum (SR) vesicles, and whole hearts were isolated from control (wildtype [WT]) or PLN knockout (pln-/-) mice. Compared to WT, pln-/- myocytes displayed enhanced resting sarcomere shortening, peak Ca2+ transient, and blunted β-adrenergic responsiveness. HNO stimulated shortening, relaxation, and Ca2+ transient in WT cardiomyocytes, and evoked positive inotropy/lusitropy in intact hearts. These changes were markedly blunted in pln-/- cells/hearts. HNO enhanced SR Ca2+ uptake in WT but not pln-/- SR-vesicles. Spectroscopic studies in insect cell microsomes expressing SERCA2a±PLN showed that HNO increased Ca2+-dependent SERCA2a conformational flexibility but only when PLN was present. In cardiomyocytes, HNO achieved this effect by stabilizing PLN in an oligomeric disulfide bond-dependent configuration, decreasing the amount of free inhibitory monomeric PLN available. Innovation: HNO-dependent redox changes in myocyte PLN oligomerization relieve PLN inhibition of SERCA2a. Conclusions: PLN plays a central role in HNO-induced enhancement of SERCA2a activity, leading to increased inotropy/lusitropy in intact myocytes and hearts. PLN remains physically associated with SERCA2a; however, less monomeric PLN is available resulting in decreased inhibition of the enzyme. These findings offer new avenues to improve Ca2+ handling in failing hearts. Antioxid. Redox Signal. 19, 1185-1197.

AB - Aims: Nitroxyl (HNO) interacts with thiols to act as a redox-sensitive modulator of protein function. It enhances sarcoplasmic reticular Ca 2+ uptake and myofilament Ca2+ sensitivity, improving cardiac contractility. This activity has led to clinical testing of HNO donors for heart failure. Here we tested whether HNO alters the inhibitory interaction between phospholamban (PLN) and the sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) in a redox-dependent manner, improving Ca 2+ handling in isolated myocytes/hearts. Results: Ventriculocytes, sarcoplasmic reticulum (SR) vesicles, and whole hearts were isolated from control (wildtype [WT]) or PLN knockout (pln-/-) mice. Compared to WT, pln-/- myocytes displayed enhanced resting sarcomere shortening, peak Ca2+ transient, and blunted β-adrenergic responsiveness. HNO stimulated shortening, relaxation, and Ca2+ transient in WT cardiomyocytes, and evoked positive inotropy/lusitropy in intact hearts. These changes were markedly blunted in pln-/- cells/hearts. HNO enhanced SR Ca2+ uptake in WT but not pln-/- SR-vesicles. Spectroscopic studies in insect cell microsomes expressing SERCA2a±PLN showed that HNO increased Ca2+-dependent SERCA2a conformational flexibility but only when PLN was present. In cardiomyocytes, HNO achieved this effect by stabilizing PLN in an oligomeric disulfide bond-dependent configuration, decreasing the amount of free inhibitory monomeric PLN available. Innovation: HNO-dependent redox changes in myocyte PLN oligomerization relieve PLN inhibition of SERCA2a. Conclusions: PLN plays a central role in HNO-induced enhancement of SERCA2a activity, leading to increased inotropy/lusitropy in intact myocytes and hearts. PLN remains physically associated with SERCA2a; however, less monomeric PLN is available resulting in decreased inhibition of the enzyme. These findings offer new avenues to improve Ca2+ handling in failing hearts. Antioxid. Redox Signal. 19, 1185-1197.

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