HUMAN ETHE1: ACTIVITY AND CATALYTIC MECHANISM

人类 ETHE1：活性和催化机制

• 批准号: 8170252
• 负责人: WOLFRAM MEYER-KLAUCKE
• 金额: $ 0.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170252
• 关键词: Acidithiobacillus; Attention; Bacteria; Binding; Cells; Computer Retrieval of Information on Scientific Projects Database; DNA Sequence; Disease; Drug Delivery Systems; Functional disorder; Funding; Genes; Genetic Transcription; Gram-Negative Bacteria; Grant; Homeostasis; Human; Human body; Institution; Ions; Iron; Lead; Life; Mammals; Measurement; Metal Binding Site; Metals; Mycobacterium tuberculosis; Organism; Oxidation-Reduction; Proteins; Regulation; Research; Research Personnel; Resources; Source; Spectrum Analysis; Time; Trace Elements; United States National Institutes of Health; Virulence Factors; absorption; base; cell injury; computerized data processing; iron metabolism; pathogen; uptake

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. Trace elements are essential for all living organisms. At the same time in cells the concentration of free metal ions is very low, because free redox-active metals can generate radicals, which can lead to cell damage. Others, such as Ca and Zn are participating in signaling processes. This implies the need for tight regulation. In mammals dysfunction of the regulation causes severe diseases. Bacteria would suffer from similar problems in case of too high or too low metal contents. Given the fact that mammalian and bacterial trace element regulation is based on different protein networks, the bacterial metal regulators have attracted considerable attention as potential drug targets. During the past years we have established our research on bacterial metal regulation. Pathogens, such as M. tuberculosis have to contend with iron sequestration in order to survive in the human body. Iron metabolism is regulated by controlling transcription of genes involved in iron uptake, transport and storage. In M. tuberculosis the ferric uptake regulator A (FurA) is activated by Fe2+ to bind specifically to its target DNA sequence thereby repressing the downstream genes. These genes include in homologous cases virulence factors and several proteins required by the bacterium for iron homeostasis. By x-ray absorption spectroscopy we will determine the metal binding sites of the protein. This allows distinguishing between two structurally and functionally distinct FurAMtb metal binding sites and provides a meticulous description plus a qualitative and quantitative characterization of them. Here we propose x-ray absorption spectroscopy measurements on the Mycobacterium tuberculosis iron uptake regulator FurA to further characterize this important gene product in the absence and presence of a new lead compound. For comparison we will include Fur_AF from Acidithiobacillus ferrooxidans, a Gram-negative bacterium living at pH2 2 in high concentrations of soluble ferrous and ferric iron.

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。 痕量元素对于所有生物体都是必不可少的。同时，在细胞中，游离金属离子的浓度非常低，因为游离氧化还原活性金属可以产生自由基，从而导致细胞损伤。其他人，例如CA和ZN正在参与信号过程。这意味着需要严格的监管。在调节的哺乳动物功能障碍中会引起严重疾病。如果金属含量过高或太低，细菌会遇到类似问题。鉴于哺乳动物和细菌痕量元件调节基于不同的蛋白质网络，细菌金属调节剂作为潜在的药物靶标引起了很大的关注。在过去的几年中，我们已经建立了对细菌金属调节的研究。 病原体，例如结核分枝杆菌必须与铁隔离以在人体中生存。铁代谢通过控制与铁摄取，传输和储存的基因的转录来调节。在结核分枝杆菌中，铁摄取调节剂A（Fura）被Fe2+激活，以特异性与其靶DNA序列结合，从而抑制下游基因。这些基因包括在同源病例中毒力因子和细菌需要用于铁稳态所需的几种蛋白质。通过X射线吸收光谱，我们将确定蛋白质的金属结合位点。 这允许区分两个结构和功能上不同的Furamtb金属结合位点，并提供细致的描述以及它们的定性和定量表征。 在这里，我们提出在结核分枝杆菌铁调节剂FURA上进行X射线吸收光谱测量，以在不存在和存在新铅化合物的情况下进一步表征该重要的基因产物。为了进行比较，我们将包括来自酸硫代杆菌的Fur_af，这是一种属于pH2 2的革兰氏阴性细菌，高浓度可溶性亚铁和铁铁。