Cardiac phosphoproteome reveals cell signaling events involved in doxorubicin cardiotoxicity
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- 作者：S¨¦verine Gratia ; Laurence Kay ; Sylvie Michell ; Michel S¨¨ve ; Uwe Schlattner ; Malgorzata Tokarska-Schlattner
- 关键词：Anthracyclines ; Cardiac cell signaling ; Cardiotoxicity ; Doxorubicin ; Phosphoproteome
- 刊名：Journal of Proteomics
- 出版年：2012
- 出版时间：3 August, 2012
- 年：2012
- 卷：75
- 期：15
- 页码：4705-4716
- 全文大小：1540 K
文摘
The successful use of anthracyclines like doxorubicin in chemotherapy is limited by their severe cardiotoxicity. Despite decades of clinical application, a satisfying description of the molecular mechanisms involved and a preventive treatment have not yet been achieved. Here we address doxorubicin-induced changes in cell signaling as a novel potential mediator of doxorubicin toxicity by applying a non-biased screen of the cardiac phosphoproteome. Two-dimensional gel electrophoresis, phosphospecific staining, quantitative image analysis, and MALDI-TOF/TOF mass spectrometry were combined to identify (de)phosphorylation events occurring in the isolated rat heart upon Langendorff-perfusion with clinically relevant (5 ¦ÌM) and supraclinical concentrations (25 ¦ÌM) of doxorubicin. This approach identified 22 proteins with a significantly changed phosphorylation status and these results were validated by immunoblotting for selected phosphosites. Overrepresentation of mitochondrial proteins (> 40 % ) identified this compartment as a prime target of doxorubicin. Identified proteins were mainly involved in energy metabolism (e.g. pyruvate dehydrogenase and acyl-CoA dehydrogenase), sarcomere structure and function (e.g. desmin) or chaperone-like activities (e.g. ¦Á-crystallin B chain and prohibitin). Changes in phosphorylation of pyruvate dehydrogenase, regulating pyruvate entry into the Krebs cycle, and desmin, maintaining myofibrillar array, are relevant for main symptoms of cardiac dysfunction related to doxorubicin treatment, namely energy imbalance and myofibrillar disorganization. This article is part of a Special Issue entitled: Translational Proteomics.