Phosphoproteome-based kinase activity profiling reveals the critical role of MAP2K2 and PLK1 in neuronal autophagy
Phosphoproteome-based kinase activity profiling reveals the critical role of MAP2K2 and PLK1 in neuronal autophagy
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DOI：
10.1080/15548627.2017.1371393
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发表时间：
2017-01-01
期刊：
影响因子：
13.3
通讯作者：
Xue, Yu
中科院分区：
文献类型：
--
作者：
Chen, Lei-Lei;Wang, Yong-Bo;Xue, Yu
最近的研究表明，宏观自噬/自噬的失调可能在神经退行性疾病的发病机制中起着核心作用，自噬的诱导通过增强其清除来保护易聚集蛋白的毒性损伤。因此，自噬已成为神经退行性疾病的一个有前途的治疗靶点。在这项研究中，定量磷酸化蛋白质组学分析与计算分析一起进行描绘的磷酸化信号网络调节的2个天然的神经保护性自噬增强剂，corynoxine（Cory）和corynoxine B（Cory B）。为了确定关键的调节因子，即蛋白激酶，我们开发了一种新的基于网络的算法，在硅片激酶组活性分析（iKAP）计算推断潜在的重要蛋白激酶的磷酸化网络。使用该算法，我们观察到Cory或Cory B可能调节几种激酶。我们预测并验证了Cory，而不是Cory B，下调了一个充分记录的自噬激酶，RPS 6 KB 1/p70 S6 K（核糖体蛋白S6激酶，多肽1）。我们还发现了2种激酶，MAP 2K 2/MEK 2（丝裂原活化蛋白激酶激酶2）和PLK 1（polo样激酶1），可能被Cory上调，而siRNA介导的Map 2k 2和Plk 1的敲低显著抑制了Cory诱导的自噬。此外，Cory通过增强自噬促进阿尔茨海默病相关APP（淀粉样β [A4]前体蛋白）和帕金森病相关SNCA/α-突触核蛋白（突触核蛋白，α）的清除，并且这些作用通过抑制MAP 2K 2和PLK 1的激酶活性而显著减弱。总的来说，我们的研究不仅开发了一种从磷酸化蛋白质组学数据中识别重要调控因子的强大方法，而且还确定了MAP 2K 2和PLK 1在神经元自噬中的重要作用。
Recent studies have demonstrated that dysregulation of macroautophagy/autophagy may play a central role in the pathogenesis of neurodegenerative disorders, and the induction of autophagy protects against the toxic insults of aggregate-prone proteins by enhancing their clearance. Thus, autophagy has become a promising therapeutic target against neurodegenerative diseases. In this study, quantitative phosphoproteomic profiling together with a computational analysis was performed to delineate the phosphorylation signaling networks regulated by 2 natural neuroprotective autophagy enhancers, corynoxine (Cory) and corynoxine B (Cory B). To identify key regulators, namely, protein kinases, we developed a novel network-based algorithm of in silico Kinome Activity Profiling (iKAP) to computationally infer potentially important protein kinases from phosphorylation networks. Using this algorithm, we observed that Cory or Cory B potentially regulated several kinases. We predicted and validated that Cory, but not Cory B, downregulated a well-documented autophagy kinase, RPS6KB1/p70S6K (ribosomal protein S6 kinase, polypeptide 1). We also discovered 2 kinases, MAP2K2/MEK2 (mitogen-activated protein kinase kinase 2) and PLK1 (polo-like kinase 1), to be potentially upregulated by Cory, whereas the siRNA-mediated knockdown of Map2k2 and Plk1 significantly inhibited Cory-induced autophagy. Furthermore, Cory promoted the clearance of Alzheimer disease-associated APP (amyloid beta [A4] precursor protein) and Parkinson disease-associated SNCA/alpha-synuclein (synuclein, alpha) by enhancing autophagy, and these effects were dramatically diminished by the inhibition of the kinase activities of MAP2K2 and PLK1. As a whole, our study not only developed a powerful method for the identification of important regulators from the phosphoproteomic data but also identified the important role of MAP2K2 and PLK1 in neuronal autophagy.