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Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart

BACKGROUND: The multifunctional Ca(2+)‐ and calmodulin‐dependent protein kinase II (CaMKII) is a crucial mediator of cardiac physiology and pathology. Increased expression and activation of CaMKII has been linked to elevated risk for arrhythmic events and is a hallmark of human heart failure. A usef...

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Autores principales: Scholten, Arjen, Preisinger, Christian, Corradini, Eleonora, Bourgonje, Vincent J., Hennrich, Marco L., van Veen, Toon A. B., Swaminathan, Paari D., Joiner, Mei‐Ling, Vos, Marc A., Anderson, Mark E., Heck, Albert J. R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Blackwell Publishing Ltd 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3828808/
https://www.ncbi.nlm.nih.gov/pubmed/23926118
http://dx.doi.org/10.1161/JAHA.113.000318
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author Scholten, Arjen
Preisinger, Christian
Corradini, Eleonora
Bourgonje, Vincent J.
Hennrich, Marco L.
van Veen, Toon A. B.
Swaminathan, Paari D.
Joiner, Mei‐Ling
Vos, Marc A.
Anderson, Mark E.
Heck, Albert J. R.
author_facet Scholten, Arjen
Preisinger, Christian
Corradini, Eleonora
Bourgonje, Vincent J.
Hennrich, Marco L.
van Veen, Toon A. B.
Swaminathan, Paari D.
Joiner, Mei‐Ling
Vos, Marc A.
Anderson, Mark E.
Heck, Albert J. R.
author_sort Scholten, Arjen
collection PubMed
description BACKGROUND: The multifunctional Ca(2+)‐ and calmodulin‐dependent protein kinase II (CaMKII) is a crucial mediator of cardiac physiology and pathology. Increased expression and activation of CaMKII has been linked to elevated risk for arrhythmic events and is a hallmark of human heart failure. A useful approach to determining CaMKII's role therein is large‐scale analysis of phosphorylation events by mass spectrometry. However, current large‐scale phosphoproteomics approaches have proved inadequate for high‐fidelity identification of kinase‐specific roles. The purpose of this study was to develop a phosphoproteomics approach to specifically identify CaMKII's downstream effects in cardiac tissue. METHODS AND RESULTS: To identify putative downstream CaMKII targets in cardiac tissue, animals with myocardial‐delimited expression of the specific peptide inhibitor of CaMKII (AC3‐I) or an inactive control (AC3‐C) were compared using quantitative phosphoproteomics. The hearts were isolated after isoproterenol injection to induce CaMKII activation downstream of β‐adrenergic receptor agonist stimulation. Enriched phosphopeptides from AC3‐I and AC3‐C mice were differentially quantified using stable isotope dimethyl labeling, strong cation exchange chromatography and high‐resolution LC‐MS/MS. Phosphorylation levels of several hundred sites could be profiled, including 39 phosphoproteins noticeably affected by AC3‐I‐mediated CaMKII inhibition. CONCLUSIONS: Our data set included known CaMKII substrates, as well as several new candidate proteins involved in functions not previously implicated in CaMKII signaling.
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spelling pubmed-38288082013-11-19 Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart Scholten, Arjen Preisinger, Christian Corradini, Eleonora Bourgonje, Vincent J. Hennrich, Marco L. van Veen, Toon A. B. Swaminathan, Paari D. Joiner, Mei‐Ling Vos, Marc A. Anderson, Mark E. Heck, Albert J. R. J Am Heart Assoc Original Research BACKGROUND: The multifunctional Ca(2+)‐ and calmodulin‐dependent protein kinase II (CaMKII) is a crucial mediator of cardiac physiology and pathology. Increased expression and activation of CaMKII has been linked to elevated risk for arrhythmic events and is a hallmark of human heart failure. A useful approach to determining CaMKII's role therein is large‐scale analysis of phosphorylation events by mass spectrometry. However, current large‐scale phosphoproteomics approaches have proved inadequate for high‐fidelity identification of kinase‐specific roles. The purpose of this study was to develop a phosphoproteomics approach to specifically identify CaMKII's downstream effects in cardiac tissue. METHODS AND RESULTS: To identify putative downstream CaMKII targets in cardiac tissue, animals with myocardial‐delimited expression of the specific peptide inhibitor of CaMKII (AC3‐I) or an inactive control (AC3‐C) were compared using quantitative phosphoproteomics. The hearts were isolated after isoproterenol injection to induce CaMKII activation downstream of β‐adrenergic receptor agonist stimulation. Enriched phosphopeptides from AC3‐I and AC3‐C mice were differentially quantified using stable isotope dimethyl labeling, strong cation exchange chromatography and high‐resolution LC‐MS/MS. Phosphorylation levels of several hundred sites could be profiled, including 39 phosphoproteins noticeably affected by AC3‐I‐mediated CaMKII inhibition. CONCLUSIONS: Our data set included known CaMKII substrates, as well as several new candidate proteins involved in functions not previously implicated in CaMKII signaling. Blackwell Publishing Ltd 2013-08-23 /pmc/articles/PMC3828808/ /pubmed/23926118 http://dx.doi.org/10.1161/JAHA.113.000318 Text en © 2013 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley-Blackwell. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article under the terms of the Creative Commons Attribution Noncommercial License, which permits use, distribution, and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Original Research
Scholten, Arjen
Preisinger, Christian
Corradini, Eleonora
Bourgonje, Vincent J.
Hennrich, Marco L.
van Veen, Toon A. B.
Swaminathan, Paari D.
Joiner, Mei‐Ling
Vos, Marc A.
Anderson, Mark E.
Heck, Albert J. R.
Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart
title Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart
title_full Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart
title_fullStr Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart
title_full_unstemmed Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart
title_short Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin‐Dependent Protein Kinase II Regulation in the Heart
title_sort phosphoproteomics study based on in vivo inhibition reveals sites of calmodulin‐dependent protein kinase ii regulation in the heart
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3828808/
https://www.ncbi.nlm.nih.gov/pubmed/23926118
http://dx.doi.org/10.1161/JAHA.113.000318
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