DYNAMICS & INTERPROTEIN INTERACTIONS IN RELEASE OF IRON IN BACTERIOFERRITIN

动力学

• 批准号: 8359665
• 负责人: Mario Rivera
• 金额: $ 6.78万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359665
• 关键词: Binding; Complex; Computing Methodologies; Coupled; Deuterium; Disease; Electron Transport; Enteral; Ferredoxin; Ferritin; Funding; Grant; Haemophilus influenzae; Homeostasis; Hydrogen; Invaded; Iron; Metabolism; Methods; Molecular; NMR Spectroscopy; National Center for Research Resources; Nutrient; Organism; Oxidoreductase; Physiological; Principal Investigator; Process; Property; Proteins; Pseudomonas aeruginosa; Research; Research Infrastructure; Resources; Respiratory Tract Infections; Shigella dysenteriae; Signal Transduction; Source; Toxic effect; United States National Institutes of Health; antimicrobial; bacterioferritin; cost; improved; pathogen; prevent; protein structure function; uptake; Binding; Complex; Computing Methodologies; Coupled; Deuterium; Disease; Electron Transport; Enteral; Ferredoxin; Ferritin; Funding; Grant; Haemophilus influenzae; Homeostasis; Hydrogen; Invaded; Iron; Metabolism; Methods; Molecular; NMR Spectroscopy; National Center for Research Resources; Nutrient; Organism; Oxidoreductase; Physiological; Principal Investigator; Process; Property; Proteins; Pseudomonas aeruginosa; Research; Research Infrastructure; Resources; Respiratory Tract Infections; Shigella dysenteriae; Signal Transduction; Source; Toxic effect; United States National Institutes of Health; antimicrobial; bacterioferritin; cost; improved; pathogen; prevent; protein structure function; uptake

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. Organisms causative of diseases such as respiratory tract infections (Haemophilus influenzae), enteric conditions (Shigella dysenteriae) and the opportunistic Pseudomonas aeruginosa have developed sophisticated mechanisms for sequestering iron from their host. This intense competition between invading pathogens and their host for the nutrient has led to the idea that new antimicrobials may target iron acquisition and homeostasis. To study this idea more closely, it is important to gain molecular-level understanding of the mechanisms by which pathogens manage iron, from acquisition and internalization to storage and utilization. Significant advances have improved our understanding of iron uptake by P. aeruginosa and many other pathogens. In comparison, little is known about the fate of internalized iron. One mechanism whereby iron toxicity is controlled is by storage of iron in ferritin and bacterioferritin, which are large proteins capable of storing up to 4,000 iron atoms in their internal cavities. Despite the importance of ferritins and bacterioferritins in regulating iron concentrations and preventing its toxic effects, little is known about the processes that deliver Fe2+ for storage or the signals that prompt its release for safe integration in metabolism. We have recently demonstrated that mobilization of Fe2+ from bacterioferritin A (BfrA) in P. aeruginosa requires electron transfer from a ferredoxin reductase (FPR). Thus the BfrA-FPR complex is an unprecedented opportunity to investigate how bacterioferritins recognize their physiological regulators and if binding modulates the dynamic properties of the bacterioferritin to facilitate iron release. To fill these gaps we plan to: (1) Investigate the dynamic properties of BfrA utilizing a strategy specifically tailored to study large proteins using hydrogen/deuterium H/D exchange coupled to NMR spectroscopy. (2) Investigate the dynamic properties of BfrA with the aid of computational methods and (3) Utilize computational and HD/NMR methods to investigate how BfrA binds to FPR and determine the effect that the inter-protein association exerts on the dynamic properties of BfrA.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 生物体的病因，例如呼吸道感染（流感嗜血杆菌），肠道疾病（Shigella dysenteriae）和机会性铜绿假单胞菌的病因已开发出复杂的机制，用于从其宿主那里隔离铁。入侵的病原体与他们的养分宿主之间的激烈竞争导致了一个想法，即新的抗菌药物可能针对铁的获取和稳态。更仔细地研究这一想法，重要的是要对病原体管理铁，从获取和内部化到存储和利用的机制进行分子级的理解。铜绿假单胞菌和许多其他病原体对我们对铁的吸收的明显进步已经提高了我们对铁的摄取的理解。相比之下，对内部铁的命运知之甚少。 控制铁毒性的一种机制是通过在铁蛋白和细菌铁蛋白中储存铁，它们的大型蛋白质能够在其内部空腔中储存多达4,000个铁原子。尽管铁蛋白和细菌铁蛋白在调节铁浓度和防止其毒性作用方面非常重要，但对提供Fe2+的储存过程或促使其在代谢中安全整合的信号的过程知之甚少。我们最近证明，铜绿假单胞菌中的细菌铁蛋白A（BFRA）动员Fe2+需要从铁氧还原还原酶（FPR）中转移电子转移。因此，BFRA-FPR复合物是研究细菌铁蛋白如何识别其生理调节剂的前所未有的机会，并且结合是否调节了细菌铁蛋白的动态特性以促进铁释放。 为了填补这些空白，我们计划：（1）研究BFRA的动态特性，该策略专门针对使用氢/氘H/D交换耦合到NMR光谱法研究了大型蛋白质。 （2）借助于计算方法研究BFRA的动态特性，（3）利用计算和HD/NMR方法来研究BFRA如何与FPR结合，并确定蛋白质间关联对BFRA动态性能的影响。