Proteomics Core

蛋白质组学核心

• 批准号: 10615815
• 负责人: ARTHUR Robert SALOMON
• 金额: $ 13.81万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-15 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10615815
• 关键词: Affinity Chromatography; Alleles; Amino Acids; Biochemical; Biological Assay; CD8B1 gene; Cell Culture Techniques; Cell Line; Cell physiology; Cells; Cellular Structures; Collaborations; Collection; Computer Analysis; Computer Models; Coupled; Custom; Data; Data Set; Detection; Effectiveness; Evaluation; Event; Feedback; Funding; Generations; Guanine Nucleotide Exchange Factors; High Pressure Liquid Chromatography; Immunology; Infrastructure; Knowledge; Label; Maps; Mass Spectrum Analysis; Mediating; Methods; Modernization; Molecular; Mus; Nature; PTPRC gene; Paper; Pathologic; Peptides; Peripheral; Phase; Phosphopeptides; Phosphorylation; Phosphorylation Site; Phosphotyrosine; Post-Translational Protein Processing; Process; Property; Protein Analysis; Proteins; Proteome; Proteomics; Publications; Publishing; Receptor Signaling; Research Personnel; Role; Sampling; Signal Pathway; Signal Transduction; Signaling Protein; Site; Source; Speed; Stable Isotope Labeling; Statistical Data Interpretation; Stimulus; Structure; System; T-Cell Receptor; T-Lymphocyte; Techniques; Therapeutic Intervention; Time; Tyrosine; Tyrosine Phosphorylation Site; Universities; ZAP-70 Gene; analog; computational suite; computerized tools; data acquisition; data analysis pipeline; design; experimental study; improved; instrument; intermolecular interaction; liquid chromatography mass spectrometry; mass spectrometer; mutant; new technology; novel; phosphoproteomics; programs; protein profiling; protein protein interaction; protein purification; recruit; response; senior faculty; src Homology Region 2 Domain; tool; transmission process

ABSTRACT – PROTEOMICS CORE Signaling networks are crucial for the orchestration of cellular functions in response to stimuli. Knowledge of the structure of these networks provides a basis for understanding the pathological consequences of their malfunction and offers opportunities for designing therapeutic interventions. The complexity of these networks and the speed with which signals are transmitted in cells makes mapping them a formidable challenge. The typical approach for elucidating the structure of cellular signaling networks involves an iterative process of creating signaling protein disruptions, domain mutants and site-directed mutants followed by characterization of each mutant through a battery of cellular activation assays. As a complementary approach, modern proteomic methods using quantitative mass spectrometry can facilitate the hypothesis-driven characterization of signaling pathways by providing a global view of cellular phosphorylation and protein-protein interactions through a variety of activation states. The Core B, Proteomics core will make cutting-edge, quantitative proteomic capabilities and computational analysis available to the investigators of this program project. The core will provide identification and relative quantitation of the protein composition and post-translational modification state of proteins using modern LC/MS techniques. This core has a strong track record of fruitful collaboration with the PI's of the program project culminating in the generation of 7 large phosphoproteomic and 8 CoIP-LCMS protein interaction datasets and publication of collaborative papers which elucidated the molecular details of how ZAP-70 is recruited to LAT and how the catalytic activity of ZAP-70 mediates basal signaling and negative feedback of T cell receptor signaling. The core has recently developed new technologies for the characterization of protein interaction networks in living cells using TurboID and the deepest possible characterization of phosphorylation networks using Src SH2 domain Superbinder and TMT BOOST channels. These newly developed methods will be leveraged to support the project PIs to determine protein-protein interactors and phosphorylation sites from T cell lines and primary mouse T cells. The core also has a suite of computational tools to provide rigorous statistical analysis of the proteomic data and to make new signaling pathway predictions.

蛋白质组学核心 信号网络对于响应刺激的细胞功能的协调至关重要。知识 这些网络的结构为理解其病理后果提供了基础。 并为设计治疗干预措施提供了机会。这些网络的复杂性 并且信号在小区中传输的速度使得映射它们成为一个艰巨的挑战。的 用于阐明蜂窝信令网络结构的典型方法包括迭代过程 产生信号蛋白破坏、结构域突变体和定点突变体， 通过一系列的细胞活化试验来检测每一个突变体。作为一种补充方法，现代 使用定量质谱的蛋白质组学方法可以促进假设驱动的表征 通过提供细胞磷酸化和蛋白质-蛋白质相互作用的全局视图， 通过各种激活状态。 核心B，蛋白质组学核心将使尖端的，定量蛋白质组学的能力和计算 分析提供给这个项目的研究人员。核心将提供识别和相对 使用现代生物技术定量蛋白质组成和蛋白质的翻译后修饰状态 LC/MS技术。这个核心有一个强大的跟踪记录，富有成效的合作与PI的计划 最终产生7个大磷酸化蛋白质组和8个CoIP-LCMS蛋白质相互作用的项目 数据集和合作论文的出版物，阐明了ZAP-70是如何被 以及ZAP-70的催化活性如何介导T细胞的基础信号传导和负反馈。 细胞受体信号传导核心最近开发了新的技术，用于蛋白质的表征 使用TurboID的活细胞中的相互作用网络和磷酸化的最深可能表征 网络使用Src SH 2域Superbinder和TMT BOOST通道。这些新开发的方法 将用于支持项目PI，以确定蛋白质-蛋白质相互作用物和磷酸化位点 来自T细胞系和原代小鼠T细胞。核心也有一套计算工具， 对蛋白质组数据进行严格的统计分析，并做出新的信号通路预测。