Structure and regulation of non-receptor tyrosine kinases

非受体酪氨酸激酶（NRTK）作为细胞信号中严格调节的整合开关发挥作用 转导在这里，我们重点介绍了三个NRTK家族的代表，它们具有不同的多域 结构：Jak家族激酶、粘着斑激酶（FAK）和Tec家族成员ITK（IL-2诱导型 酪氨酸激酶）。我们的长期目标是阐明NRTKs在结构上的自调节机制， 水平，了解这些调节机制是如何在癌症中被破坏的，并利用结构上的见解， 有助于发现新的抑制剂。与我们的合作者，我们正在结合结构生物学的工具， 生物化学，细胞生物学和化学以统一的方式推进这些目标。我们提出三个具体的 目标。首先，我们将发现Jak激酶如何识别它们的同源细胞因子受体，以及它们是如何被激活的。 由其组成域之间的相互作用调节。我们已经结晶了一个N末端片段， Jak 2的这一部分，并且还制备了Jak 2的这一部分与促红细胞生成素的胞质尾的复合物 用于结构分析的受体。我们对Jak调节的研究建立在我们最近确定的一个 接头/假激酶调节模块，并通过阐明基本上全长Jak 2的结构， 我们将解释这个模块如何控制相邻激酶结构域的活性，以及这种控制是如何 在骨髓增生性肿瘤中被V617 F突变破坏。其次，我们将从结构上确定如何 粘着斑激酶（FAK）被PI（4，5）P2激活。这一目标建立在我们确定 FAK在其自身抑制状态，以及我们发现它结合并被PI（4，5）P2激活。三是 阐明Itk的SH 3-SH 2-激酶片段的结构，以了解其调节，并促进 抑制剂发现基于我们的结构见解，我们正在开发针对Jak 3的不可逆抑制剂。 其可用于治疗自身免疫和炎性疾病。

Non-receptor tyrosine kinases (NRTKs) function as tightly regulated, integrating switches in cellular signal transduction. Here we focus on representatives of three NRTK families, which have distinct multi-domain architectures: Jak-family kinases, Focal Adhesion Kinase (FAK), and the Tec-family member ITK (Il-2 inducible tyrosine kinase). Our longterm goals are to elucidate autoregulatory mechanisms of NRTKs at a structural level, to understand how these regulatory mechanisms are disrupted in cancer, and to use structural insights to facilitate discovery of novel inhibitors. With our collaborators, we are combining the tools of structural biology, biochemistry, cell biology, and chemistry in a unified way to advance these goals. We propose three Specific Aims. First, we will discover how Jak kinases recognize their cognate cytokine receptors and how they are regulated by interactions among their constituent domains. We have crystallized an Nterminal fragment of Jak2, and have also prepared a complex of this portion of Jak2 with the cytoplasmic tail of the erythropoietin receptor for structural analysis. Our studies of Jak regulation build on our recently determined structure of a linker/pseudokinase regulatory module, and through elucidation of the structure of essentially full-length Jak2, we will explain how the this module controls the activity of the adjacent kinase domain, and how this control is disrupted by the V617F mutation in myeloproliferative neoplasms. Second, we will determine structurally how focal adhesion kinase (FAK) is activated by PI(4,5)P2. This aim builds on our determination of the structure of FAK in its autoinhibited state, and our discovery that it binds and is activated by the PI(4,5)P2. Third, we will elucidate the structure an SH3-SH2-kinase fragment of Itk in order to understand its regulation and to facilitate inhibitor discovery. Based on our structural insights, we are developing irreversible inhibitors specific for Jak3 that may be useful in treatment of autoimmune and inflammatory disorders.