PROTEOMIC ANALYSIS OF BIOMARKERS AND MECHANISMS OF TOXIC METAL STRESS

生物标志物的蛋白质组学分析和有毒金属应激机制

• 批准号: 7723537
• 负责人: Susan Mary Miller
• 金额: $ 0.58万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723537
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Biological Markers; Cells; Chimera organism; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Databases; Drug Metabolic Detoxication; Escherichia coli; Exposure to; Funding; Grant; Homology Modeling; Imagery; Institution; Mass Spectrum Analysis; Mercury; Modification; Operon; Pathway interactions; Peptides; Plasmids; Post-Translational Protein Processing; Proteins; Proteomics; Reactive Oxygen Species; Research; Research Personnel; Resources; Source; Stress; Structure; Sulfhydryl Compounds; United States National Institutes of Health; Work; adduct; base; dithiol; protein structure; software development; tool; toxic metal

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. In this project, we are using a mass spectrometry proteomics approach to identify the cellular effects of exposure to Hg(II) and organomercurials. The initial project involves studies of E. coli cells that do or do not harbor a plasmid that carries the mer operon which encodes a bacterial mercury detoxification pathway. We have hypothesized that there may be two types of protein modifications resulting from exposure to the Hg compounds, (1) formation of Hg(II) and RHg(I) adducts with protein thiols, and (2) oxidiative modifications that may result if initial targets of Hg interaction stimulate formation of reactive oxygen species. One fairly straightforward aspect of work in my lab is to use a structure-based approach to build predictions of expected Hg(II)-dithiol and RHg(I)-thiol complexes of proteins to augment the databases used to identify modified peptides found in the mass spec analyses. The second aspect that has proved more important is to develop software tools to automate the analysis of the mass spec data to look for Hg-containing adducts and identify the peptides/proteins. While the second aspect does not really require use of Chimera, the first aspect does and further analysis of identified targets is anticipated to involve visualization of protein structures or homology models.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 在这个项目中，我们使用质谱蛋白质组学方法来识别暴露于Hg（II）和有机核的细胞影响。 最初的项目涉及研究或不携带质粒的大肠杆菌细胞，该质粒带有编码细菌汞排毒途径的MER操纵子。 我们假设可能有两种类型的蛋白质修饰，导致暴露于Hg化合物，（1）Hg（II）和RHG（I）与蛋白硫醇的形成，以及（2）氧化性修饰，如果Hg相互作用的初始靶标刺激了反应性氧气的形成，则可能导致氧化性修饰。 我实验室中工作的一个相当直接的方面是使用基于结构的方法来构建对蛋白质的预期Hg（II） - 二硫醇和RHG（I） - 硫醇复合物的预测，以增强用于识别质量规格分析中发现的修改肽的数据库。 事实证明，更重要的第二个方面是开发软件工具以自动化质量规格数据的分析以寻找含HG的加合物并识别肽/蛋白质。 虽然第二个方面并不需要真正使用嵌合体，但第一个方面确实需要使用，并且预计确定的靶标的进一步分析涉及蛋白质结构或同源性模型的可视化。