Core B: Qunatitative Analysis and Proteomics Core to Detect S-Nitrosylation

核心 B：检测 S-亚硝基化的定量分析和蛋白质组学核心

• 批准号: 7559780
• 负责人: Zezong Gu
• 金额: $ 12.64万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7559780
• 关键词: Biological Assay; Biological Markers; Biological Models; Biotin; Brain; Cell model; Cells; Chemicals; Code; Complex; Condition; Cysteine; Detection; Dissociation; Electron Transport; In Vitro; Individual; Ions; Isotopes; Label; Lysine; Mass Spectrum Analysis; Mediating; Methods; Mission; Mitochondria; Modification; N-terminal; Nature; Neurons; Paper; Peptides; Phase; Protein Disulfide Isomerase; Proteins; Proteomics; Publishing; Reaction; Resolution; Science; Screening procedure; Solid; Sulfhydryl Compounds; Sulfonic Acids; Techniques; Technology; Testing; high throughput screening; in vivo; inhibitor/antagonist; neuroprotection; oxidation; parkin gene/protein; prevent; protein function; small molecule; small molecule libraries; synuclein

The mission of this Quantitative Analysis and Proteomics Core (QAPC) will be to provide comprehensive proteomic approaches for the individual projects of this NIEHS Center to detect and characterize PD-related proteins, especially with regard to S-nitrosylation (reaction of the NO group with critical cysteine thiol to regulate protein function) and further oxidation reactions. Core B will support the efforts of Project 1, Project 2, and Project 3 by analysis and quantification of PDrelated proteins and their possible S-nitrosylation (including SNO-parkin, SNO-DJ-1, and SNOprotein disulfide isomerase [SNO-PDI]) using multiple chemical and proteomic approaches. Core B will also support the efforts of Project 4 by detecting possible biomarkers represented by these SNO-proteins in a-synuclein-over-expressing or mitochondrial complex I inhibitor-treated in hESCderived neuronal cell models. This Core will also interact with Core E (High-throughput Chemical Library Screening) by providing DAN assays that detect S-nitrosylated proteins in 384- and 1536- well multiwell formats in order to screen for small molecules that prevent S-nitrosylation of PDrelated proteins such as parkin, DJ-1 and PDI. These small molecules will then be tested for neuroprotection in PD model systems in Projects 2, 3, and 4.

这个定量分析和蛋白质组学核心(QAPC)的任务是提供 NIEHS中心单个项目的综合蛋白质组学方法以检测和 帕金森病相关蛋白的特征，特别是关于S-亚硝化(NO基团的反应 用关键的半胱氨酸硫醇调节蛋白质功能)和进一步的氧化反应。核心B将 通过分析和量化与PD相关的项目，支持项目1、项目2和项目3的工作 蛋白质及其可能的S亚硝化(包括SNO-parkin、SNO-DJ-1和SNO蛋白 二硫键异构酶[SNO-PDI])使用多种化学和蛋白质组学方法。核心B 还将通过检测这些代表的可能的生物标志物来支持项目4的工作 α-突触核蛋白过度表达或线粒体复合体I抑制剂处理hESCs中的SNO蛋白 神经细胞模型。该核心还将与核心E(高通量化学品 文库筛选)，通过提供检测S亚硝化蛋白的DAN试验 多井格式化以筛选阻止PDR的S-亚硝化的小分子 Parkin、DJ-1和PDI等蛋白质。然后这些小分子将被测试 项目2、3和4中帕金森病模型系统的神经保护。