Proteomic Profiling of Cardiomyocyte-Specific Cathepsin A Overexpression Links Cathepsin A to the Oxidative Stress Response.
Proteomic Profiling of Cardiomyocyte-Specific Cathepsin A Overexpression Links Cathepsin A to the Oxidative Stress Response.
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DOI：
10.1021/acs.jproteome.6b00413
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发表时间：
2016-07
影响因子：
4.4
通讯作者：
Agnese Petrera;U. Kern;D. Linz;Alejandro Gomez-Auli;M. Hohl;Johann Gassenhuber;T. Sadowski;O. Schilling
中科院分区：
文献类型：
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作者：
Agnese Petrera;U. Kern;D. Linz;Alejandro Gomez-Auli;M. Hohl;Johann Gassenhuber;T. Sadowski;O. Schilling
组织蛋白酶A(Cthepsin A，CTSA)是一种存在于细胞表面、分泌于细胞外的溶酶体羧肽酶。此外，CTSA还与β-半乳糖苷酶和神经氨酸酶1结合，以保护它们免受降解。CTSA作为治疗心肌肥厚和心力衰竭的药物靶点已引起人们的关注。在这篇论文中，我们在心肌细胞特异性的人CTSA过度表达的小鼠模型中，结合同位素二甲基二甲基标记策略，研究了CTSA对小鼠心肌蛋白质组的影响。我们在三个生物复制中的每个复制中确定了多达2000个蛋白质。通过对微阵列数据的线性模型的统计分析(LIMMA)发现>；300显著影响蛋白质(适度的p值≤0.01)，从而使ctSA成为心脏蛋白质组的关键调节因子。CTSA通过上调几种蛋白水解酶，如组织蛋白酶B、组织蛋白酶D和组织蛋白酶Z来强烈破坏蛋白分解系统的平衡，同时下调许多蛋白水解酶抑制剂。此外，心肌细胞特异性的人CTSA过表达显著降低了许多抗氧化应激蛋白的水平，即过氧化还蛋白和蛋白脱甘氨酸DJ-1。在体外，利用培养的大鼠心肌细胞，CTSA的异位过表达导致了活性氧的积累。总的来说，我们的蛋白质组学和功能数据加强了CTSA与细胞氧化应激反应之间的联系。
Cathepsin A (CTSA) is a lysosomal carboxypeptidase present at the cell surface and secreted outside the cell. Additionally, CTSA binds to β-galactosidase and neuraminidase 1 to protect them from degradation. CTSA has gained attention as a drug target for the treatment of cardiac hypertrophy and heart failure. Here, we investigated the impact of CTSA on the murine cardiac proteome in a mouse model of cardiomyocyte-specific human CTSA overexpression using liquid chromatography-tandem mass spectrometry in conjunction with an isotopic dimethyl labeling strategy. We identified up to 2000 proteins in each of three biological replicates. Statistical analysis by linear models for microarray data (limma) found >300 significantly affected proteins (moderated p-value ≤0.01), thus establishing CTSA as a key modulator of the cardiac proteome. CTSA strongly impaired the balance of the proteolytic system by upregulating several proteases such as cathepsin B, cathepsin D, and cathepsin Z while down-regulating numerous protease inhibitors. Moreover, cardiomyocyte-specific human CTSA overexpression strongly reduced the levels of numerous antioxidative stress proteins, i.e., peroxiredoxins and protein deglycase DJ-1. In vitro, using cultured rat cardiomyocytes, ectopic overexpression of CTSA resulted in accumulation of reactive oxygen species. Collectively, our proteomic and functional data strengthen an association of CTSA with the cellular oxidative stress response.