STRUCTURE/FUNCTION AND PROTEIN-PROTEIN INTERACTION ANALYSIS IN HG DETOXIFICATION

HG 解毒中的结构/功能和蛋白质-蛋白质相互作用分析

• 批准号: 8363729
• 负责人: Susan Mary Miller
• 金额: $ 0.15万
• 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: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363729
• 关键词: Bacterial Proteins; Complex; Drug Metabolic Detoxication; Enzymes; Funding; Goals; Grant; Individual; Ions; Kinetics; Mass Spectrum Analysis; Membrane Transport Proteins; Mercury; Metals; Mutate; National Center for Research Resources; Pathway interactions; Principal Investigator; Property; Protein Analysis; Proteins; Proteolysis; Research; Research Infrastructure; Resources; Role; Site-Directed Mutagenesis; Source; Structure; United States National Institutes of Health; chemical property; cost; oxidation; physical property; protein protein interaction; protein structure

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 investigating the role of protein-protein interactions in the mechanism of metal ion transfers between the key proteins of the bacterial detoxification pathway for inorganic mercury and organomercurials and elucidating the chemical and physical properties embedded in the protein structures that facilitate the directed transfer of Hg(II) to the essential enzyme for reductive detoxification. The structures of four components of the pathway have been determined and the structure of an integral membrane transport protein is being pursued to facilitate structure/function analysis of the proteins. Together with the structural information we use site-directed mutagenesis to evaluate the effects of altered structural features on the individual properties and functions of the several proteins and on their interactions. The goal of the studies is to reveal the features of the proteins that are essential from an evolutionary perspective for this detoxification pathway to be effective. Mass spec has proved invaluable in verifying oxidation and proteolysis problems with several of our mutated enzymes and in verifying formation of metal ion complexes that we have used in our structural and kinetic studies.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 在这个项目中，我们正在研究蛋白质-蛋白质相互作用在无机汞细菌解毒途径的关键蛋白质和有机汞之间的金属离子转移机制中的作用，并阐明蛋白质结构中嵌入的化学和物理性质，促进汞(II)直接转移到还原解毒的关键酶。已经确定了该途径的四个组成部分的结构，并正在寻找一个完整的膜运输蛋白的结构，以便于对这些蛋白的结构/功能进行分析。与结构信息一起，我们使用定点突变来评估结构特征改变对几个蛋白质的单独属性和功能以及它们之间相互作用的影响。这项研究的目的是从进化的角度揭示这种解毒途径有效所必需的蛋白质的特征。事实证明，质谱仪在验证我们的几种突变酶的氧化和蛋白质分解问题以及验证我们在结构和动力学研究中使用的金属离子络合物的形成方面具有极高的价值。