New Proteomic Technologies for the Analysis of Tyrosine Kinase Signaling Pathways

用于分析酪氨酸激酶信号通路的蛋白质组学新技术

• 批准号: 8325721
• 负责人: W. Andy Tao
• 金额: $ 34.49万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8325721
• 关键词: Affinity; Biochemical; Biological Models; Breast Cancer Cell; Cancer Biology; Cell Proliferation; Cells; Characteristics; Chemicals; Clinical Data; Complex Mixtures; Coupled; Data; Development; Disease; Event; Goals; Growth; Immune; Individual; Knowledge; Lead; Link; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Medicine; Methodology; Methods; Modeling; Modification; Molecular; Monitor; Neoplasm Metastasis; Outcome; Pathway interactions; Phosphopeptides; Phosphoproteins; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiological; Play; Polymers; Property; Protein Tyrosine Kinase; Proteins; Proteomics; Protocols documentation; Research; Research Personnel; Role; Series; Signal Pathway; Signal Transduction; Site; Squamous cell carcinoma; System; Techniques; Technology; Testing; Therapeutic; Tumor Suppressor Proteins; Tyrosine Phosphorylation; Tyrosine Phosphorylation Site; base; cancer cell; cancer prevention; cancer type; cell growth; cell growth regulation; cell motility; cell type; design; human SYK protein; improved; in vivo; innovation; interest; leukemia/lymphoma; malignant breast neoplasm; malignant phenotype; melanoma; neoplastic cell; novel; outcome forecast; overexpression; pre-clinical; public health relevance; response; tool

DESCRIPTION (provided by applicant): A key objective of phosphoproteomic analyses is to obtain an in-depth understanding of protein phosphorylation and signaling networks in cells. However, despite the great progress that has been made over the past few years, our ability to analyze such signaling pathways is still greatly limited by a number of serious technical challenges. The long term goal is to develop novel proteomics strategies valuable for understanding on the molecular level the role of phosphorylation in regulating cell proliferation. The objective in this application is to devise and optimize effective and efficient methodologies and strategies for the analyses of tyrosine kinase phosphoproteomes, using a dual functional tyrosine kinase Syk as the model system. This approach will utilize functional, soluble polymers for the efficient and inclusive isolation of phosphopeptides coupled with an integrated chemical, mass spectrometric, and computational strategy for the identification of specific tyrosine kinase substrates and for quantitative phosphoproteomics. We will focus on Syk-dependent signaling and establish protocols that will ultimately provide a powerful method to dissect any signaling pathway regulated by protein-tyrosine phosphorylation. Guided by strong preliminary data, this objective will be achieved by pursuing three specific aims: 1) Develop a proteomic platform for the analysis of protein phosphorylation. We will primarily develop, characterize and optimize functionalized soluble polymers designed for the affinity and chemical isolation of phosphopeptides; 2) Identification of specific tyrosine-kinase substrates and their phosphorylation sites; and 3) In-depth analyses of tyrosine kinase phosphoproteomes in several cancer types. We will develop and apply our quantitative proteomic technologies for the identification and quantification of tyrosine-phosphorylation sites in several cancer types. This project is innovative because it will develop a series of novel analytical techniques to directly identify novel tyrosine kinase substrates and phosphorylation sites that will provide investigators with a significantly expanded capability to dissect signaling pathways. The developed techniques will provide detailed knowledge at the molecular level on precisely how tyrosine kinases are involved in cell signaling under different physiological conditions through dynamic changes in protein phosphorylation. It is also expected that, by identifying a kinase's direct substrates, the proposed research will provide a powerful tool to conveniently map kinase networks. PUBLIC HEALTH RELEVANCE: The proposed studies to develop a series of analytical techniques have potential applicability for investigators in the molecular signaling field. The tools will allow them to examine important roles of tyrosine kinase in multiple signaling pathways. The proposed research will provide unique strategies and materials that will strengthen the important contribution that proteomics can make to our understanding of cancer biology.

描述（由申请人提供）：磷酸化蛋白质组学分析的一个关键目标是深入了解细胞中的蛋白质磷酸化和信号网络。然而，尽管在过去几年中取得了很大的进展，但我们分析这些信号通路的能力仍然受到许多严重技术挑战的极大限制。长期目标是开发新的蛋白质组学策略，在分子水平上理解磷酸化在调节细胞增殖中的作用。本申请的目的是设计和优化有效和高效的方法和策略，用于分析酪氨酸激酶磷酸化蛋白质组，使用双功能酪氨酸激酶Syk作为模型系统。这种方法将利用功能性的可溶性聚合物，用于磷酸肽的有效和包容性分离，并结合集成的化学，质谱和计算策略，用于识别特定的酪氨酸激酶底物和定量磷酸蛋白质组学。我们将专注于Syk依赖的信号转导和建立协议，最终将提供一个强大的方法来剖析任何由蛋白质酪氨酸磷酸化调节的信号通路。在强大的初步数据的指导下，这一目标将通过追求三个具体目标来实现：1）开发一个蛋白质组学平台，用于分析蛋白质磷酸化。我们将主要开发，表征和优化功能化可溶性聚合物，用于磷酸肽的亲和和化学分离; 2）识别特定的酪氨酸激酶底物及其磷酸化位点; 3）深入分析几种癌症类型中的酪氨酸激酶磷酸化蛋白质组。我们将开发和应用我们的定量蛋白质组学技术，用于识别和定量几种癌症类型中的酪氨酸磷酸化位点。该项目是创新的，因为它将开发一系列新的分析技术，以直接识别新的酪氨酸激酶底物和磷酸化位点，这将为研究人员提供一个显着扩展的能力，剖析信号通路。开发的技术将提供详细的知识，在分子水平上，正是如何酪氨酸激酶参与细胞信号在不同的生理条件下，通过蛋白质磷酸化的动态变化。我们还希望，通过识别激酶的直接底物，所提出的研究将提供一个强大的工具，方便地映射激酶网络。 公共卫生相关性：所提出的研究，以开发一系列的分析技术具有潜在的适用性，研究人员在分子信号领域。这些工具将使他们能够研究酪氨酸激酶在多种信号通路中的重要作用。这项研究将提供独特的策略和材料，加强蛋白质组学对我们理解癌症生物学的重要贡献。