Proteomics Core

蛋白质组学核心--项目摘要 SC Cobre中蛋白质组学核心在氧化剂、氧化还原平衡和胁迫信号方面的广泛目标 (Redox Cobre)是提供最先进的、基于质谱学(MS)的蛋白质组能力，以使 阐明疾病背后的氧化还原信号机制、细胞和药物的治疗效果 治疗抵抗力。核心提供专业知识、技术援助、仪器和解释 定量蛋白质组学实验的分析结果和氧化还原敏感型后基因组的表征 翻译修饰语。在第一和第二阶段，获得Orbitrap Elite ETD MS和Orbitrap聚变 Lumos ETD/UVPD MS Nano-LC-MS/MS系统(NIH S10 OD010731和S10 OD025126，PI：Lauren Ball) 允许使用互补多肽片段方法(ETD)检测具有挑战性的修饰 和EThcD)以及Cobre所需的高级、定量蛋白质组工作流程的开发 调查人员。在第三阶段，核心将进一步发展和更新这些能力，以反映快速 氧化还原蛋白质组学相关仪器、软件和方法学的进展。此外，在 响应Cobre附属研究人员的需求，使用MALDI提供蛋白质组组织成像功能- 将提供成像MS(IMS)来识别酶消化肽在体内的空间分布 玻片上的组织生物标记物用于发现可能的生物标志物和疾病的潜在机制 进步。LC-MS检测疾病特异性蛋白和氧化还原敏感的翻译后修饰 MS/MS和IMS提出了对有针对性的蛋白质组能力的需求，以验证这些发现并进一步 推进翻译研究。提供这种蛋白质组学技术的补充将建立在成功的基础上 对核心进行定位，以建立一个全面、独立的资源。核心已经利用 NIGMS在Cobre计划第一阶段和第二阶段期间所做的先前投资，OD和NIGMS 共享仪器资助，NCI癌症中心支持资助，以及长期、强大的机构 致力于发展成为对Redox Cobre需求至关重要的成功机构资源 调查人员。目前，蛋白质组学核心有19个与Cobre有关的用户，他们是15个活跃的PI NIH奖项依赖于LC-MS/MS能力，其中五个奖项也依赖于IMS能力。

Proteomics Core – Project Summary The broad objective of the Proteomics Core in the SC COBRE in Oxidants, Redox Balance, and Stress Signaling (Redox COBRE) is to provide state-of-the-art, mass spectrometry (MS)-based proteomic capabilities to enable elucidation of redox signaling mechanisms underlying disease, therapeutic efficacy of cells and drugs, and therapeutic resistance. The Core provides expertise, technical assistance, instrumentation, and interpretation of analytical results for quantitative proteomic experiments and the characterization of redox-sensitive post- translational modifications. During Phase I and II, acquisition of the Orbitrap Elite ETD MS and Orbitrap Fusion Lumos ETD/UVPD MS nano-LC-MS/MS systems (NIH S10 OD010731 and S10 OD025126, PI: Lauren Ball) permitted detection of challenging modifications using complementary peptide fragmentation approaches (ETD and EThcD) and the development of advanced, quantitative proteomic workflows as needed by COBRE investigators. During Phase III, the Core will further develop and update these capabilities to reflect rapid advances in instrumentation, software, and methodologies relevant to redox proteomics. Additionally, in response to the needs of COBRE-affiliated investigators, proteomic tissue imaging capabilities using MALDI- Imaging MS (IMS) will be provided to discern the spatial distribution of enzymatically-digested peptides within tissue biospecimens on slides for the discovery of putative biomarkers and mechanisms underlying disease progression. Detection of disease-specific proteins and redox-sensitive post-translational modifications by LC- MS/MS and IMS has raised the need for targeted proteomic capabilities to verify these findings and further advance translational studies. Providing this complement of proteomic technologies will build on the successes of the Core and position the Core to establish a comprehensive, independent resource. The Core has leveraged previous investments made by the NIGMS during Phase I and II of the COBRE program, the OD and NIGMS for Shared Instrumentation Grants, the NCI Cancer Center Support Grant, and long-standing, strong institutional commitment to develop into a successful institutional resource critical to the needs of Redox COBRE investigators. At present there are 19 COBRE-affiliated users of the Proteomics Core that are PIs of 15 active NIH awards that rely on the LC-MS/MS capabilities, five of these awards also depend on IMS capabilities.