Proteomic Core

蛋白质组核心

• 批准号: 8882844
• 负责人: AKHILESH PANDEY
• 金额: $ 23.49万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-30 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8882844
• 关键词: Acetylation; Address; Affinity; Amino Acids; Automobile Driving; Basic Science; Bioinformatics; Cations; Cell Culture Techniques; Chemicals; Clinical; Clinical Research; Communities; Coupling; Data Set; Digestion; Disease; Disease model; Fractionation; Goals; Human; Investigation; Label; Liquid Chromatography; Mammals; Mass Spectrum Analysis; Methodology; Methods; Molecular; Molecular Profiling; Parkinson Disease; Pathogenesis; Pathology; Patients; Peptides; Phase; Phosphorylation; Play; Post-Translational Protein Processing; Preparation; Proteins; Proteome; Proteomics; Research; Research Project Grants; Resolution; Resource Sharing; Resources; Role; Sampling; Signal Transduction; Signaling Protein; Stable Isotope Labeling; TNFRSF5 gene; Technology; Validation; base; computerized tools; innovation; instrument; mouse model; multiple reaction monitoring; novel; protein expression; tool

SUMMARY The Proteomics Core will employ cutting-edge proteomics technologies to help address mechanistic questions proposed by the four JHM Udall Center research projects regarding the pathogenesis of Parkinson's disease (PD). The methodologies available through the Proteomics Core provide efficient and sensitive strategies to identify and quantify PD-associated changes in the samples obtained from both disease models and human patients. Thus, we anticipate that proteomic findings generated by the Proteomics Core will help elucidate specific protein-based cellular mechanisms that correlate with PD pathogenesis, identifying novel pathogenic proteins, signaling nodes and protein PTMs with strong association to PD. To this end, the Proteomics Core will provide innovative and traditional mass spectrometry (MS)-based approaches to facilitate the identification and quantification of proteins and their major post-translational modifications (PTMs) including phosphorylation, ubiquitylation and acetylation in the context of diseases. The Core has a wide breadth of expertise in discovery as well as targeted quantitative proteomics methods and strategies. This includes stable isotope labeling by amino acids in cell culture (SILAC) labeling, stable isotope labeling in mammals (SILAM), tandem mass tags (TMT) labeling, filter-aided sample preparation (FASP) based protein extraction and digestion, peptide fractionation methods such as basic reverse-phase liquid chromatography (bRPLC) and strong cation exchange (SCX), chemical and affinity-based PTM-enrichment methods, high-resolution high- accuracy mass spectrometry analysis for global proteomic profiling, targeted quantitation strategies including Multiple Reaction Monitoring (MRM), and bioinformatics and computational tools for the interpretation of even large-scale datasets. The combination of these high-end technologies offers powerful tools to address the proposed research questions of the JHM Udall Center. The overall goal of the Proteomics Core (Core D) is to use state-of-the-art mass spectrometry-based proteomics to facilitate the discovery and validation of protein molecules implicated in PD pathogenesis, providing support to all proposed basic science projects within this Udall Center and to the broader Morris K. Udall scientific community involved in PD research.

概括 蛋白质组学核心将采用尖端蛋白质组学技术来帮助解决机制问题 由 JHM Udall 中心的四个关于帕金森病发病机制的研究项目提出 （PD）。通过蛋白质组学核心可用的方法提供了高效且灵敏的策略 识别并量化从疾病模型和人类获得的样本中与 PD 相关的变化 患者。因此，我们预计蛋白质组学核心产生的蛋白质组学发现将有助于阐明 与 PD 发病机制相关的基于特定蛋白质的细胞机制，识别新的致病因素 与 PD 密切相关的蛋白质、信号转导节点和蛋白质 PTM。为此，蛋白质组学核心 将提供创新和传统的基于质谱（MS）的方法来促进识别 蛋白质及其主要翻译后修饰 (PTM) 的定量和定量，包括 疾病背景下的磷酸化、泛素化和乙酰化。核心具有广泛的范围 发现方面的专业知识以及有针对性的定量蛋白质组学方法和策略。这包括稳定的 细胞培养中的氨基酸同位素标记 (SILAC) 标记、哺乳动物中的稳定同位素标记 (SILAM)、 串联质量标签 (TMT) 标记、基于过滤辅助样品制备 (FASP) 的蛋白质提取和 消化、肽分级分离方法，例如基本反相液相色谱 (bRPLC) 和 强阳离子交换 (SCX)、基于化学和亲和力的 PTM 富集方法、高分辨率高分辨率 用于全球蛋白质组分析的准确质谱分析，有针对性的定量策略，包括 多重反应监测 (MRM) 以及用于解释甚至解释的生物信息学和计算工具 大规模数据集。这些高端技术的结合提供了强大的工具来解决 提出了 JHM 尤德尔中心的研究问题。蛋白质组学核心（核心 D）的总体目标是 使用最先进的基于质谱的蛋白质组学来促进蛋白质的发现和验证 与 PD 发病机制有关的分子，为该领域内所有拟议的基础科学项目提供支持 尤德尔中心以及参与 PD 研究的更广泛的 Morris K. Udall 科学界。