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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在这个项目中，我们使用质谱蛋白质组学方法来确定暴露于 Hg(II) 和有机汞对细胞的影响。 最初的项目涉及对大肠杆菌细胞的研究，这些细胞是否含有携带 mer 操纵子的质粒，该操纵子编码细菌汞解毒途径。 我们假设，暴露于汞化合物可能会导致两种类型的蛋白质修饰，(1) 与蛋白质硫醇形成汞 (II) 和 RHg(I) 加合物，以及 (2) 如果汞相互作用的初始目标刺激活性氧的形成，则可能导致氧化修饰。 我实验室工作的一个相当简单的方面是使用基于结构的方法来构建蛋白质的预期 Hg(II)-二硫醇和 RHg(I)-硫醇复合物的预测，以增强用于识别质谱分析中发现的修饰肽的数据库。 事实证明更重要的第二个方面是开发软件工具来自动分析质谱数据，以寻找含汞加合物并识别肽/蛋白质。 虽然第二个方面并不真正需要使用嵌合体，但第一个方面需要使用嵌合体，并且对已识别靶标的进一步分析预计将涉及蛋白质结构或同源模型的可视化。