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: 8363617
• 负责人: Susan Mary Miller
• 金额: $ 1.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 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 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363617
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Biological Markers; Cells; Chimera organism; Data; Databases; Drug Metabolic Detoxication; Escherichia coli; Exposure to; Funding; Grant; Homology Modeling; Imagery; Informatics; Mass Spectrum Analysis; Mercury; Modification; National Center for Research Resources; Operon; Pathway interactions; Peptides; Plasmids; Post-Translational Protein Processing; Principal Investigator; Proteins; Proteomics; Reactive Oxygen Species; Research; Research Infrastructure; Resources; Software Tools; Source; Stress; Structure; Sulfhydryl Compounds; United States National Institutes of Health; Work; adduct; base; biocomputing; cost; dithiol; protein complex; protein structure; software development; toxic metal

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 在这个项目中，我们正在使用质谱蛋白质组学的方法来确定暴露于汞（II）和有机汞的细胞效应。 本研究的第一个项目是对E.大肠杆菌细胞中携带或不携带携带编码细菌汞解毒途径的mer操纵子的质粒。 我们假设，可能有两种类型的蛋白质修饰导致暴露于汞化合物，（1）形成汞（II）和RHg（I）与蛋白质硫醇的加合物，（2）氧化修饰，可能导致如果汞相互作用的初始目标刺激活性氧的形成。 在我的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建立预期的Hg（II）-二硫醇和RHg（I）-硫醇蛋白质复合物的预测，以增加用于识别质谱分析中发现的修饰肽的数据库。 已被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鉴定肽/蛋白质。 虽然第二方面并不真正需要使用嵌合体，但第一方面需要，并且预期对鉴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。