STRUCTURAL INVESTIGATION OF FOCAL ADHESION MOLECULES

描述(申请人提供)：通过连接细胞外基质(ECM)蛋白和肌动蛋白细胞骨架，整合素蛋白调节细胞的增殖、迁移和存活。粘着斑激酶(FAK)在整合素信号转导中起着关键作用。FAK对局部粘连的激活和定位是整合素与ECM蛋白结合的结果。激活的FAK将信号从整合素传递到不同通路的下游组件。然而，FAK的不适当激活和不适当的FAK调节信号与癌症和其他人类疾病有关。因此，我们的长期目标是研究FAK调控的信号通路的调控和靶向相互作用的结构和分子机制。这一目标将通过使用生物物理方法来实现。 研究表明，FAK的C末端焦点黏附靶向(FAT)结构域是FAK靶向焦点黏附的必要条件和充分条件，这是FAK信号转导的关键步骤。根据在本申请中描述的初步发现，假设FAT结构域通过与FAK的N端FERM结构域相互作用来调节FAK的激活。此前，首席研究人员通过核磁共振波谱确定脂肪结构域的溶液结构。在拟议的研究中，这种方法将被用来确定脂肪结构域与其结合伙伴--巴西林、他林和FAK的FERM结构域之间相互作用的化学性质。 此外，还将评估FAK、巴西林和他林之间的复杂分子相互作用。 这些研究将在原子水平上提供有关脂肪结构域及其相关蛋白质的结构和功能信息。此外，设计用作脂肪结构域抑制剂的配体将进行测试，以确定它们是否在开发专门干扰FAK介导的导致癌症的异常信号的治疗方面发挥主导作用。

DESCRIPTION (provided by applicant): By linking extracellular matrix (ECM) proteins to the actin cytoskeleton, integrin proteins regulate cell proliferation, migration, and survival. A key player in integrin signaling is focal adhesion kinase (FAK). Activation and localization of FAK to focal adhesions results from the binding of integrins to ECM proteins. Activated FAK relays signal from integrins to downstream components of different pathways. However, inappropriate activation of FAK and inappropriate FAK-regulated signaling has been implicated in cancer and other human diseases. Therefore, the long-term objective is to investigate structures and molecular mechanisms underlying regulatory and targeting interactions of FAK-regulated signaling pathways. This objective will be achieved through the use of biophysical methods. Studies showed that FAK's C-terminal focal adhesion targeting (FAT) domain is necessary and sufficient for the targeting of FAK to focal adhesions, a crucial step in FAK signaling. On the basis of preliminary findings described in this application, it is hypothesized that the FAT domain regulates FAK activation by interacting with FAK's N-terminal FERM domain. Previously, the principal investigator determined the solution structure of the FAT domain by NMR spectroscopy. In proposed studies, this method will be used to determine the chemical nature of interactions between the FAT domain and its binding partners: paxillin, talin, and FAK's FERM domain. Furthermore, the complex molecular interplay among FAK, paxillin, and talin will be evaluated. These studies will provide structural and functional information at the atomic level about the FAT domain and its associated proteins. Also, ligands designed to serve as FAT domain inhibitors, will be tested to determine whether they provide a lead in developing therapies that interfere specifically with abnormal FAK-mediated signaling that contributes to cancer.