Journal article
Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin-Dependent Protein Kinase II Regulation in the Heart
Journal of the American Heart Association, Vol.2(4), pp.e000318-n/a
08/01/2013
DOI: 10.1161/JAHA.113.000318
PMCID: PMC3828808
PMID: 23926118
Abstract
Background-The multifunctional Ca2+- 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 beta-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.
Details
- Title: Subtitle
- Phosphoproteomics Study Based on In Vivo Inhibition Reveals Sites of Calmodulin-Dependent Protein Kinase II Regulation in the Heart
- Creators
- Arjen Scholten - Utrecht UniversityChristian Preisinger - Utrecht UniversityEleonora Corradini - Utrecht UniversityVincent J. Bourgonje - University Medical Center UtrechtMarco L. Hennrich - Utrecht UniversityToon A. B. van Veen - University Medical Center UtrechtPaari D. Swaminathan - Roy J. and Lucille A. Carver College of MedicineMei-Ling Joiner - Roy J. and Lucille A. Carver College of MedicineMarc A. Vos - University Medical Center UtrechtMark E. Anderson - University of IowaAlbert J. R. Heck - Utrecht University
- Resource Type
- Journal article
- Publication Details
- Journal of the American Heart Association, Vol.2(4), pp.e000318-n/a
- Publisher
- Wiley
- DOI
- 10.1161/JAHA.113.000318
- PMID
- 23926118
- PMCID
- PMC3828808
- ISSN
- 2047-9980
- eISSN
- 2047-9980
- Number of pages
- 11
- Grant note
- Netherlands Proteomics Centre in the Netherlands Genomics Initiative R01HL079031 / NATIONAL HEART, LUNG, AND BLOOD INSTITUTE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Heart Lung & Blood Institute (NHLBI) R01HL70250; R01HL079031; R01HL113001; R01HL096652 / NIH; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA Utrecht University's Focus Massa program Fondation Leducq: the Alliance for CaMKII Signaling in Heart Disease; Leducq Foundation
- Language
- English
- Date published
- 08/01/2013
- Academic Unit
- Biology; Internal Medicine
- Record Identifier
- 9984366377502771
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