Probing tyrosine phosphatase structure and function

探究酪氨酸磷酸酶的结构和功能

• 批准号: 10201679
• 负责人: Neel H Shah
• 金额: $ 40.01万
• 依托单位: COLUMBIA UNIV NEW YORK MORNINGSIDE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10201679
• 关键词: Animals; Area; Biochemical; Biological; Biological Process; Biology; Cell physiology; Cells; Development; Disease; Drug Targeting; Drug resistance; Enzymes; Goals; Human; Knowledge; Label; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Methods; Molecular; Mutate; Mutation; PTPN11 gene; Phenotype; Phosphoric Monoester Hydrolases; Play; Point Mutation; Post-Translational Protein Processing; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; Regulation; Research; Role; Signal Transduction; Structural Biochemistry; Structure; T-Lymphocyte; Techniques; Therapeutic; Tyrosine; Tyrosine Phosphorylation; Work; biochemical tools; cell type; experimental study; human disease; immune activation; interest; mutation screening; novel strategies; preference; resistance mutation; targeted cancer therapy; targeted treatment; tool

PROJECT SUMMARY The enzymatic modification of proteins through tyrosine phosphorylation is a common mechanism for relaying information in animal cells. Tyrosine kinases act in a signal-responsive manner to phosphorylate specific proteins at tyrosine residues, and the opposing tyrosine phosphatases dephosphorylate proteins to dynamically regulate signals. Tyrosine phosphorylation is essential to many biological processes in healthy cells, and the dysregulation of tyrosine phosphorylation is a common feature of many diseases, most notably cancers. Over the past few decades, we have developed an extensive understanding of tyrosine kinase function and regulation, but our knowledge of tyrosine phosphatases lags behind. This disparity is partly due to the fact that it is easier to develop drugs that target tyrosine kinases than tyrosine phosphatases, and the therapeutic potential of tyrosine kinases has motivated the development of robust tools to study their structure, biochemistry, and biology. The overarching goals of my lab are to understand, at the molecular level, how tyrosine phosphatases select substrate proteins to dephosphorylate, how they are regulated through dynamic changes in their structure, and how they contribute to healthy and disease-associated signaling. Over the next five years, my group will devise new techniques to study tyrosine phosphatases. We are currently developing a high-throughput biochemical platform to rapidly identify and compare the substrate sequence preferences of tyrosine phosphatases. These analyses will be conducted in parallel with proximity-labeling experiments in live cells to tag the interaction partners of tyrosine phosphatases for identification by mass spectrometry. Together, these approaches will allow us to map the substrates of tyrosine phosphatases and help define the signaling roles of these enzymes. We are also developing methods to rapidly characterize the functional effects of all possible point mutations in a tyrosine phosphatase. These mutational screens will allow us to identify new modes of regulation, pinpoint the functional consequences of disease-associated mutations, and map likely drug-resistance mutations that may arise to phosphatase-targeted cancer therapies. We are broadly interested in two areas of signaling biology: diseases where tyrosine phosphatases are mutated and/or dysregulated, and the activation of immune T cells. As we develop new biochemical tools, we will initially apply these tools to the tyrosine phosphatase SHP2, which plays a causal role in several congenital diseases and cancers, and is also critical to normal signaling in many cell types, including T cells. Our work will clarify the signaling functions of SHP2, connect known mutations to specific phenotypes, and help guide the development of SHP2-targeted therapies. In the long-term, we will apply our novel approaches to other tyrosine phosphatases.

项目概要 通过酪氨酸磷酸化对蛋白质进行酶促修饰是一种常见机制 用于在动物细胞中传递信息。酪氨酸激酶以信号响应方式发挥作用 在酪氨酸残基处磷酸化特定蛋白质，以及相反的酪氨酸磷酸酶 使蛋白质去磷酸化以动态调节信号。酪氨酸磷酸化对许多人来说至关重要 健康细胞中的生物过程，酪氨酸磷酸化失调是一种常见的现象 许多疾病的特征，尤其是癌症。在过去的几十年里，我们开发了 对酪氨酸激酶功能和调节的广泛了解，但我们对酪氨酸的了解 磷酸酶落后。这种差异的部分原因是开发药物更容易 靶向酪氨酸激酶而不是酪氨酸磷酸酶，并且酪氨酸激酶的治疗潜力 激励了开发强大的工具来研究其结构、生物化学和生物学。 我实验室的首要目标是在分子水平上了解酪氨酸如何 磷酸酶选择底物蛋白进行去磷酸化，它们如何通过动态调节 它们结构的变化，以及它们如何促进健康和疾病相关的信号传导。超过 未来五年，我的小组将设计新技术来研究酪氨酸磷酸酶。我们目前 开发高通量生化平台以快速识别和比较底物 酪氨酸磷酸酶的序列偏好。这些分析将与 在活细胞中进行邻近标记实验，以标记酪氨酸磷酸酶的相互作用伙伴 通过质谱鉴定。总之，这些方法将使我们能够绘制基质图 酪氨酸磷酸酶并帮助定义这些酶的信号传导作用。我们也在开发 快速表征酪氨酸中所有可能的点突变的功能影响的方法 磷酸酶。这些突变筛选将使我们能够识别新的调控模式，查明 疾病相关突变的功能后果，并绘制可能的耐药突变图谱 可能会出现磷酸酶靶向癌症疗法。 我们对信号生物学的两个领域广泛感兴趣：酪氨酸参与的疾病 磷酸酶突变和/或失调，以及免疫 T 细胞的激活。随着我们的发展 新的生化工具，我们首先将这些工具应用到酪氨酸磷酸酶SHP2上，它起着 在多种先天性疾病和癌症中起因果作用，并且对许多正常信号传导也至关重要 细胞类型，包括 T 细胞。我们的工作将阐明SHP2的信号功能，连接已知的 特定表型的突变，并有助于指导 SHP2 靶向疗法的开发。在 从长远来看，我们将把我们的新方法应用于其他酪氨酸磷酸酶。