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

基本信息

批准号：
7986538
负责人：
W. Andy Tao
金额：
$ 30.13万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
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/7986538
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

项目摘要

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）：磷蛋白质组学分析的关键目标是深入了解细胞中蛋白质磷酸化和信号网络。但是，尽管在过去几年中取得了巨大进展，但我们分析此类信号通路的能力仍然受到许多严重的技术挑战的限制。长期目标是制定新颖的蛋白质组学策略对于理解分子水平的磷酸化在调节细胞增殖中的作用而有价值。本应用的目的是使用双重功能性酪氨酸激酶SYK作为模型系统来设计和优化有效，有效的方法和策略，以分析酪氨酸激酶磷酸蛋白酶的分析。这种方法将利用功能性的可溶性聚合物来有效且包容性的磷酸肽与综合化学，质谱和计算策略的磷酸肽，以鉴定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依赖性信号传导，并建立方案，该方案最终将提供一种强大的方法来剖定由蛋白质酪氨酸磷酸化调节的任何信号通路。在强大的初步数据的指导下，将通过追求三个特定目的来实现这一目标：1）开发一个蛋白质组学平台来分析蛋白质磷酸化。我们将主要开发，表征和优化旨在用于磷酸肽亲和力和化学分离的官能化的可溶性聚合物。 2）鉴定特定的酪氨酸激酶底物及其磷酸化位点； and 3) In-depth analyses of tyrosine kinase phosphoproteomes in several cancer types.我们将开发并应用我们的定量蛋白质组学技术来鉴定和定量几种癌症类型的酪氨酸磷酸化位点。该项目具有创新性，因为它将开发一系列新型的分析技术，以直接识别新型的酪氨酸激酶底物和磷酸化位点，这些酪氨酸激酶底物和磷酸化位点将为研究人员提供显着扩展的能力以剖析信号通路。开发的技术将通过蛋白质磷酸化的动态变化在不同生理条件下与酪氨酸激酶在不同生理条件下的细胞信号传导的准确方式提供详细的知识。还可以预期，通过识别激酶的直接底物，拟议的研究将为方便地映射激酶网络提供强大的工具。公共卫生相关性：开发一系列分析技术的拟议研究对分子信号领域中的研究人员具有潜在的适用性。这些工具将使他们能够检查酪氨酸激酶在多种信号通路中的重要作用。拟议的研究将提供独特的策略和材料，以加强蛋白质组学对我们对癌症生物学理解的重要贡献。