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: 8292292
• 负责人: Zezong Gu
• 金额: $ 12.38万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8292292
• 关键词: Biological Assay; Biological Markers; Biological Models; Biotin; Brain; Cell model; Cells; Chemicals; Code; Complex; Cysteine; Detection; Dissociation; Electron Transport; Grant; In Vitro; Individual; Ions; Isotopes; Label; Lysine; Mass Spectrum Analysis; Mediating; Methods; Mission; Mitochondria; Modification; N-terminal; National Institute of Environmental Health Sciences; Nature; Neurons; PINK1 gene; Paper; Parkinson Disease; Peptides; Phase; Protein Disulfide Isomerase; Protein S; Proteins; Proteomics; Publishing; Reaction; Resolution; S-Nitrosothiols; SKIL gene; Science; Screening procedure; Solid; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Sulfhydryl Compounds; Sulfonic Acids; Techniques; Technology; Testing; alpha synuclein; high throughput screening; in vivo; inhibitor/antagonist; neuroprotection; oxidation; parkin gene/protein; prevent; protein complex; protein function; small molecule; small molecule libraries

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.

该定量分析和蛋白质组学核心（QAPC）的使命将提供 全面的蛋白质组学方法，为这个NIEHS中心的个别项目，以检测和 表征PD相关蛋白，特别是关于S-亚硝基化（NO基团的反应 具有关键的半胱氨酸巯基以调节蛋白质功能）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-亚硝基化（包括SNO-parkin、SNO-DJ-1和SNOprotein 二硫键异构酶[SNO-PDI]）。核心B 还将通过检测这些生物标志物来支持项目4的工作。 α-突触核蛋白过表达或线粒体复合物I转运蛋白处理的hESC源性细胞中的SNO蛋白 神经元细胞模型。该核心还将与核心E（高通量化学分析）相互作用。 文库筛选），通过提供检测384-和1536-中S-亚硝基化蛋白的DAN测定， 为了筛选阻止PD相关的S-亚硝基化的小分子， Parkin、DJ-1和PDI等蛋白质。这些小分子将被检测 项目2、3和4中PD模型系统的神经保护。