Coordination Chemistry of Microbial Iron Transport Compounds

项目摘要/摘要 美国每年报告的细菌性败血症病例接近50万例，其中约三分之一 这些病例是致命的。铁是微生物生长中的一种限制性营养物质；细菌主要通过 铁载体、低分子量络合剂的生产，具有高的铁亲和力和选择性。这个 铁的有效性对于确定入侵病原体的毒力是至关重要的。最成功的 人类病原体，如炭疽芽孢杆菌，设计出精心设计的多方面策略，以确保他们的 铁供应。本项目旨在了解铁载体运输系统：1)从结构层面， 研究铁结合的热力学和动力学，2)在系统的水平上，遵循认识和 将这些铁载体运输到细菌中，3)到环境水平，探索环境如何， 如温度、宿主免疫系统、其他细菌的存在，甚至暴露在阳光下，都会影响 细菌的生长。我们在实验室中拥有独特的设备来进行这一系列的研究并 追求以下具体目标：1.理解结构和功能之间的关系 铁载体。2.研究铁载体介导的铁在革兰氏阳性菌中的转运。3.探索 微生物铁运输的铁载体穿梭机制的范围和功能。4.进一步 描述人类免疫系统蛋白质识别铁载体的机制(铁载体) 以及这种免疫反应的选择性如何被最危险的细菌病原体利用。 为了达到这些目标，铁载体的特性，如热力学稳定性和还原潜力 铁载体铁络合物、它们的铁结合动力学和亲脂性将被确定为靶标。 铁载体，并通过构建合成的铁载体类似物和配位类似物，我们将 探索铁载体的功能。几乎所有已知的关于细菌铁载体介导的铁 运输在革兰氏阴性菌中；革兰氏阳性菌现在是我们的目标，因为这群 生物体包括许多重要的人类病原体。我们已经进行了适当的合作来确定 补体芽孢杆菌的膜相关蛋白受体的晶体结构 我们在这个家庭里的学习。我们关于铁载体穿梭机制的第一份报告显示，金属交换 在所研究的革兰氏阴性细菌中，两个铁载体之间对于铁的运输是必不可少的。我们计划 看看这种机制在细菌属中的分布有多广泛；我们现在提出一个扩展的 结合使用同位素标记的天然铁载体的实验方法。我们最近做了 开始形成对我们所说的“铁载体隐形”的理解：通过 铁载体的结构修饰。通过使用合成类似物和细菌铁载体 分离株，以及标记底物和突变蛋白，我们打算描述其选择性和 铁胆碱的生理过程。

Project Summary/Abstract Nearly half a million cases of bacterial sepses are reported annually in the USA and approximately one third of the cases are fatal. Iron is a limiting nutrient in microbial growth; bacteria primarily obtain iron through production of siderophores, low molecular weight chelating agents with high ferric affinity and selectivity. The availability of iron is essential in determining the virulence of an invading pathogen. The most successful human pathogens, such as Bacillus anthracis, devise elaborate, multifaceted strategies to ensure their iron supply. This project seeks to understand siderophore transport systems: 1) from a structural level, studying the thermodynamics and kinetics of iron binding, 2) to a systemic level, following the recognition and transport of these siderophores into the bacteria, 3) to an environmental level, exploring how the surroundings, such as temperature, host immune system, presence of other bacteria and even exposure to light, affect the growth of the bacteria. We are uniquely equipped in our laboratory to carry out this range of studies and to pursue the following specific aims: 1. To understand the relationship between structure and function of siderophores. 2. To characterize siderophore-mediated iron transport in Gram-positive bacteria. 3. To explore the scope and functioning of the siderophore shuttle mechanism of microbial iron transport. 4. To further describe the mechanism of recognition of siderophores by proteins of the human immune system (siderocalin) and how the selectivity of this immune response is exploited by the most dangerous bacterial pathogens. To meet these Aims siderophore features such as thermodynamic stability and reduction potential of siderophore ferric complex, their kinetics of iron binding, and lipophilicity, will be determined for targeted siderophores and through the construction of synthetic siderophore analogs and coordination analogs we will explore siderophore function. Almost everything that is known about bacterial siderophore-mediated iron transport is in Gram-negative bacteria; Gram-positive bacteria are now our target, since this group of organisms includes many important human pathogens. We have in place collaborations to determine the crystallographic structures of membrane-associated protein receptors of Bacillus species that will complement our studies in this family. Our first report of the siderophore shuttle mechanism showed that metal exchange between two siderophores was essential for iron transport in the Gram-negative bacteria studied. We plan to see how widely distributed is this mechanism is among genera of bacteria; we now propose an extended experimental approach that incorporates the use of isotopically labeled natural siderophores. We have recently begun to develop an understanding of what we call "siderophore stealth": the evasion of siderocalin binding by structural modification of the siderophore. Through the use of synthetic analogs and bacterial siderophore isolates, as well as labeled substrates and mutant proteins, we intend to describe the selectivity and physiological course of siderocalin.