Increased level of phosphorylated desmin and its degradation products in heart failure.
Increased level of phosphorylated desmin and its degradation products in heart failure.
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DOI：
10.1016/j.bbrep.2016.02.014
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发表时间：
2016-07
影响因子：
2.7
通讯作者：
Pinet F
中科院分区：
文献类型：
--
作者：
Bouvet M;Dubois-Deruy E;Alayi TD;Mulder P;El Amranii M;Beseme O;Amouyel P;Richard V;Tomavo S;Pinet F
尽管已经确定了几个风险因素，如梗死面积，但心力衰竭（HF）的进展/严重程度在临床实践中仍然难以预测。使用实验性大鼠缺血性HF模型和磷酸化蛋白质组学技术，我们发现HF大鼠左心室（LV）磷酸化结蛋白水平增加。本工作的目的是评估结蛋白是否是HF的循环生物标志物或仅是HF的组织生物标志物。我们使用了几种抗体，以检测结蛋白，其蛋白水解片段和其磷酸化形式在LV和血浆中的蛋白质印迹，磷酸盐亲和电泳，质谱和免疫荧光。血浆用组合肽配体库处理或去除白蛋白和免疫球蛋白以增加检测的灵敏度。我们发现了2倍增加的丝氨酸-结蛋白磷酸化在HF大鼠的LV，主要是在不溶性部分，这表明结蛋白聚集体的形成。在HF大鼠的LV中也检测到结蛋白裂解产物，表明结蛋白磷酸化的增加导致对蛋白水解活性的更敏感性，可能由钙蛋白酶活性介导。天然结蛋白及其降解产物在大鼠、小鼠或人血浆中检测不到。这些数据表明，丝氨酸磷酸化形式的结蛋白及其降解产物，而不是结蛋白本身，作为HF的组织而不是循环生物标志物的潜力。结蛋白主要表达于大鼠左心室不溶性部分。在实验性心力衰竭中，结蛋白在丝氨酸中高度磷酸化。血浆中未检出结蛋白及其降解产物。
Although several risk factors such as infarct size have been identified, the progression/severity of heart failure (HF) remains difficult to predict in clinical practice. Using an experimental rat model of ischemic HF and phosphoproteomic technology, we found an increased level of phosphorylated desmin in the left ventricle (LV) of HF-rats. The purpose of the present work is to assess whether desmin is a circulating or only a tissue biomarker of HF. We used several antibodies in order to detect desmin, its proteolytic fragments and its phosphorylated form in LV and plasma by western blot, phosphate affinity electrophoresis, mass spectrometry and immunofluorescence. Plasma was treated with combinatorial peptide ligand library or depleted for albumin and immunoglobulins to increase the sensitivity of detection. We found a 2-fold increased serine-desmin phosphorylation in the LV of HF-rats, mainly in the insoluble fraction, suggesting the formation of desmin aggregates. Desmin cleavage products were also detected in the LV of HF rats, indicating that the increased phosphorylation of desmin results in more susceptibility to proteolytic activity, likely mediated by calpain activity. The native desmin and its degradation products were undetectable in the plasma of rat, mouse or human. These data suggest the potential of serine-phosphorylated form of desmin and its degradation products, but not of desmin itself, as tissue but not circulating biomarkers of HF. Desmin is mainly expressed in insoluble fraction of rat left ventricle. In experimental heart failure, desmin is highly phosphorylated in serine. Desmin and its degradation products are not detected in plasma.