Dynamics and Topology of Phosphotyrosine-SH2 Interaction Networks

描述（由申请方提供）：与酪氨酸激酶连接的细胞表面受体控制许多重要的细胞活性，包括增殖、分化和运动。酪氨酸激酶信号传导失调是许多人类癌症的共同特征，因此酪氨酸激酶及其下游效应物是开发用于治疗癌症的新药的靶标。然而，为了充分利用这些有前途的新疗法，我们需要了解酪氨酸激酶信号网络如何在系统水平上处理信息。虽然在开发描述酪氨酸激酶信号网络的定量模型方面取得了相当大的进展，但由于缺乏有关酪氨酸磷酸化的变化如何与含有模块化磷酸酪氨酸结合结构域的下游效应物偶联的定量信息，这些努力受到严重阻碍。这个合作项目的目标是利用新的实验方法来直接解决这一知识差距。具体来说，我们将使用SH2分析，磷酸化蛋白质组学的方法，是高度互补的质谱为基础的方法，定量受体酪氨酸激酶激活后的特定效应蛋白的结合位点的动态变化。对不同受体和不同细胞类型的反应将进行比较，使系统水平的行为与生物输出相关。我们还将使用单分子成像方法来监测细胞内环境中特定效应物与受体的偶联。这些研究将提供前所未有的深入了解受体信号传导复合物的相互作用动力学，这将使酪氨酸激酶信号传导的更强大和准确的模型。相关性（参见说明）：来自具有酪氨酸激酶活性的受体的信号传导在许多人类疾病，特别是癌症中起重要作用。精确描述这些受体所使用的信号传导机制的定量计算机模型将在设计癌症新疗法和决定哪些患者将从这些疗法中受益最多（个体化药物）方面非常有用。拟议的研究使用创新的实验方法来揭示建立更强大和更准确的模型所需的新机制见解。

DESCRIPTION (provided by applicant): Cell surface receptors linked to tyrosine kinases control a host of important cellular activities, including proliferation, differentiation, and motility. Disregulated tyrosine kinase signaling is a common feature of many human cancers, thus tyrosine kinases and their downstream effectors are targets for the development of new drugs for the treatment of cancer. In order to take full advantage of such promising new therapies, however, we need an understanding of how tyrosine kinase signaling networks process information on a systems level. While considerable progress has been made in developing quantitative models describing tyrosine kinase signaling networks, these efforts are severely hampered by a lack of quantitative information on how changes in tyrosine phosphorylation are coupled to their downstream effectors containing modular phosphotyrosine binding domains. The goals of this collaborative project are to take advantage of new experimental approaches to address this gap in knowledge directly. Specifically, we will use SH2 profiling, a phosphoproteomic approach that is highly complementary to mass spectrometry-based methods, to quantify dynamic changes in binding sites for specific effector proteins upon receptor tyrosine kinase activation. Responses to different receptors and in different cell types will be compared, allowing systems-level behavior to be correlated with biological outputs. We will also use single-molecule imaging methods to monitor the coupling of specific effectors to receptors in the intracellular environment. These studies will afford unprecedented insight into the interaction dynamics of receptor signaling complexes that will enable much more powerful and accurate models of tyrosine kinase signaling. RELEVANCE (See instructions): Signaling from receptors with tyrosine kinase activity plays an important role in a number of human diseases, in particular cancer. Quantitative computer-based models that accurately describe the signaling mechanism used by these receptors will be very useful in designing new therapies for cancer and in deciding which patients will benefit most from those therapies (individualized medicine). The proposed studies use innovative experimental approaches to reveal new mechanistic insights necessary for building more powerful and accurate models.