Proteomic Core

总结 蛋白质组学核心将采用尖端的蛋白质组学技术来帮助解决机制问题 由JHM Udall中心的四个研究项目提出，关于帕金森病的发病机制， （PD）。通过蛋白质组学核心可用的方法提供了有效和敏感的策略， 鉴定和量化从疾病模型和人类获得的样品中的PD相关变化 患者因此，我们预计蛋白质组学核心产生的蛋白质组学研究结果将有助于阐明 与PD发病机制相关的基于特定蛋白质的细胞机制， 蛋白质、信号传导节点和蛋白质PTM与PD有很强的关联。为此，Proteomics Core 将提供创新的和传统的质谱（MS）为基础的方法，以促进识别 蛋白质及其主要翻译后修饰（PTM）的定量，包括 磷酸化、泛素化和乙酰化。核心具有广泛的 在发现以及有针对性的定量蛋白质组学方法和策略的专业知识。这包括稳定 细胞培养物中氨基酸同位素标记（SILAC）标记，哺乳动物中稳定同位素标记（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、肽分级分离方法如碱性反相液相色谱法（bRPLC）和 强阳离子交换（SCX），化学和亲和PTM富集方法，高分辨率高， 用于全球蛋白质组学分析的准确性质谱分析，靶向定量策略，包括 多反应监测（MRM），生物信息学和计算工具，甚至解释 大规模数据集。这些高端技术的结合为解决 JHM Udall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 参与PD发病机制的分子，为所有提出的基础科学项目提供支持。 尤德尔中心和更广泛的莫里斯K。参与PD研究的Udall科学界。

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.