Shared Resources Core 2: Quantitative Proteomics Core

共享资源核心 2：定量蛋白质组学核心

• 批准号: 10024849
• 负责人: Benjamin A Garcia
• 金额: $ 27.95万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-09 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10024849
• 关键词: Affect; Antibodies; Archives; Biochemical; Biology; Cell Line; Cells; Chromatin; Chromatin Structure; Chromatography; Communities; Computing Methodologies; Enhancement Technology; Enzymes; Epigenetic Process; Epitopes; Formalin; Gene Mutation; Genes; Genetic Transcription; Goals; Histones; Human; In Vitro; Injections; Isotope Labeling; Isotopes; Malignant Neoplasms; Mammalian Cell; Mass Spectrum Analysis; Measurement; Methods; Modeling; Modification; Mutation; Oncogenic; Paraffin Embedding; Pennsylvania; Post Translational Modification Analysis; Post-Translational Modification Site; Post-Translational Protein Processing; Process; Program Research Project Grants; Protein Analysis; Protein Conformation; Proteins; Proteome; Proteomics; Research; Research Personnel; Resource Sharing; Role; Sampling; Signal Pathway; Signaling Protein; Structure; Techniques; Technology; Therapeutic; Time; Tissues; Tumor Tissue; Universities; Work; base; combinatorial; design; experimental study; histone modification; human disease; improved; in vivo; member; new technology; next generation; non-histone protein; novel; polypeptide; programs; protein expression; protein protein interaction; sortase; structural biology; technology development; tumor; tumorigenesis

Project Summary The Quantitative Proteomics core led by Dr. Benjamin Garcia will provide cutting-edge mass spectrometry (MS) based proteomics technologies to the P01 Project team members to help elucidate biochemical mechanisms involved in histone and histone-like mutations in human cancers. These MS approaches will include quantitative experiments to understand protein expression and post-translational modifications (PTMs) on histone and non-histone proteins from both cultured and primary cell lines, tissue/tumors and formalin fixed paraffin embedded (FFPE) archived samples. Although most researchers in the epigenetics and chromatin biology fields utilize antibody based methods for histone modification characterization, antibodies have many technical issues (such as epitope occlusion), which confound the analyses. Mass spectrometry therefore provides a more unbiased and complementary approach, that is also more sensitive and accurate for protein PTM analysis. The Garcia Lab has for over a decade now developed methods for histone PTM analyses. Here in this propose we plan to continue to expand our proteomics toolbox to allow P01 team members to be able to characterize 500 histone PTM sites in rapid fashion, perform quantitative analyses of histone combinatorial modifications, and determine which oncogenic histone mutations affect chromatin structure. Additionally, we will explore the downstream protein signaling pathways that are altered in epigenetically driven human cancers, as transcriptional profiles do not always provide the most accurate pictures of protein expression. !

项目摘要 由本杰明·加西亚博士领导的定量蛋白质组学核心将提供尖端的质谱分析 (MS)基于蛋白质组学技术的P01项目团队成员，以帮助阐明生物化学 人类癌症中组蛋白和组蛋白样突变的机制。这些MS方法将 包括定量实验，以了解蛋白质表达和翻译后修饰（PTM） 对来自培养细胞系和原代细胞系、组织/肿瘤和福尔马林固定的组蛋白和非组蛋白蛋白的影响 石蜡包埋（FFPE）存档样本。尽管大多数表观遗传学和染色质领域的研究人员 生物学领域利用基于抗体的方法进行组蛋白修饰表征，抗体具有许多 技术问题（如表位封闭），这混淆了分析。因此， 提供了一种更加公正和互补的方法，对蛋白质也更加敏感和准确。 PTM分析。加西亚实验室已经开发了十多年的组蛋白PTM分析方法。这里 在此建议中，我们计划继续扩展我们蛋白质组学工具箱，使P01团队成员能够 以快速方式表征500个组蛋白PTM位点，对组蛋白组合进行定量分析， 修饰，并确定哪些致癌组蛋白突变影响染色质结构。另外我们 将探索在表观遗传驱动的人类中改变的下游蛋白质信号通路。 癌症，因为转录谱并不总是提供最准确的蛋白质表达图片。 !