System-Level Analysis of the Effects of RPTPs on Cellular Signaling Networks.
System-Level Analysis of the Effects of RPTPs on Cellular Signaling Networks.
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RPTP 对细胞信号网络影响的系统级分析。
DOI：
10.1007/978-1-0716-3569-8_10
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发表时间：
2024
期刊：
影响因子：
--
通讯作者：
White,ForestM
中科院分区：
文献类型：
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作者：
Gerritsen,Jacqueline;Rizzo,Sophie;Thévenin,Damien;White,ForestM
酪氨酸磷酸化调节受体酪氨酸激酶（RTK）激活下游的信号网络活性。受体蛋白酪氨酸磷酸酶（RPTP）用于使RTK及其近端衔接蛋白去磷酸化，从而用于调节RTK活性。虽然RPTP的一般功能已经很好地理解，但每种RPTP的直接和间接底物的特征很差。在这里，我们描述了一种方法，定量磷酸酪氨酸磷酸化蛋白质组学，使特定的磷酸化位点的磷酸化水平的表达和活性的一个给定的RPTP改变的识别。在概念验证应用中，我们使用这种方法来突出显示PTPRJ（也称为DEP 1）的几个直接或间接底物磷酸化位点，并在EGF刺激的细胞中，与活性增加的PTPRJ突变形式相比，在野生型PTPRJ的背景下显示其定量磷酸化。该方法一般适用于定义不同生理条件下细胞或组织中每种RPTP的信号网络效应。
Tyrosine phosphorylation regulates signaling network activity downstream of receptor tyrosine kinase (RTK) activation. Receptor protein tyrosine phosphatases (RPTPs) serve to dephosphorylate RTKs and their proximal adaptor proteins, thus serving to modulate RTK activity. While the general function of RPTPs is well understood, the direct and indirect substrates for each RPTP are poorly characterized. Here we describe a method, quantitative phosphotyrosine phosphoproteomics, that enables the identification of specific phosphorylation sites whose phosphorylation levels are altered by the expression and activity of a given RPTP. In a proof-of-concept application, we use this method to highlight several direct or indirect substrate phosphorylation sites for PTPRJ, also known as DEP1, and show their quantitative phosphorylation in the context of wild-type PTPRJ compared to a mutant form of PTPRJ with increased activity, in EGF-stimulated cells. This method is generally applicable to define the signaling network effects of each RPTP in cells or tissues under different physiological conditions.