PROTEIN-NO INTERACTIONS PROBED BY CONVENTIONAL AND TIME-RESOLVED X-RAY CRYSTA

通过传统和时间分辨 X 射线晶体探测蛋白质-无相互作用

• 批准号: 8363698
• 负责人: MARIUS SCHMIDT
• 金额: $ 3.65万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363698
• 关键词: Bacteria; Cattle; Funding; Grant; Liver; Medicine; Methodology; Methods; Monitor; Mycobacterium tuberculosis; National Center for Research Resources; Nitric Oxide; Parasites; Phase; Physiological; Principal Investigator; Propionibacterium; Proteins; Public Health; Research; Research Infrastructure; Resources; Roentgen Rays; Role; Signal Transduction; Source; Structure; Superoxide Dismutase; System; Testing; Time; United States National Institutes of Health; Virus; X-Ray Crystallography; antimicrobial; antimicrobial drug; catalase; cost; fungus; structural biology; time use; tool

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. Since the late 80s nitric oxide has been recognized to have many important physiological roles. For example NO has important signaling functions and is a potent antimicrobial agent active against a wide range of bacteria viruses fungi and even parasites. Nitric oxides physiological functions almost invariably involve specific interactions with proteins; much progress has been made in identifying the mechanisms of such protein-NO interactions but many questions remain. Herein we propose to develop broadly applicable methodologies for photochemically generating free NO within a protein crystal so that subsequent interactions of NO with the protein can be monitored in real time using time-resolved protein X-ray crystallography. Such methodologies will be invaluable tools for studying the detailed mechanisms of NO signaling and antimicrobial phenomena. The project will be divided into 2 phases. In phase 1 various methods for NO photogeneration will be tested on crystals of bovine liver catalase and superoxide dismutase (SOD) from Propionibacterium shermanii. Both are readily available proteins whose crystal structures are known and that interact strongly with NO. In the second phase of the project the best NO photogenerating systems will be used to investigate NO interaction with oxygenated HbN from M. tuberculosis. HbN is a protein that is thought to protect M. tuberculosis from the bodys defenses and allow TB to persist for years in a latent state. The aim of this phase will be to show that our method can be applied to a system of considerable importance in medicine and public health.

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。 自80年代后期以来，一氧化氮被认为具有许多重要的生理作用。例如，NO具有重要的信号传导功能，并且是一种有效的抗菌剂，以对广泛的细菌病毒真菌甚至寄生虫进行活性。一氧化氮生理功能几乎总是涉及与蛋白质的特定相互作用。在确定这种蛋白质相互作用的机制方面取得了很多进展，但仍然存在许多问题。本文中，我们建议开发广泛适用的方法，用于在蛋白质晶体内进行光化学生成无的NO，以便可以使用时间分辨蛋白X射线晶体学实时监测NO与蛋白质的随后相互作用。这种方法将是研究无信号传导和抗菌现象的详细机制的宝贵工具。该项目将分为两个阶段。在第1阶段，将对来自叶杆菌的牛肝酶晶体和超氧化物歧化酶（SOD）的晶体进行各种无光生的方法。两者都是易于获得的蛋白质，其晶体结构是已知的，并且与NO相互作用。在项目的第二阶段中，最佳的无光生成系统将用于研究与结核分枝杆菌的含氧HBN相互作用。 HBN是一种蛋白质，被认为可以保护结核分枝杆菌免受身体防御的影响，并使结核病在潜在状态下持续多年。此阶段的目的是表明我们的方法可以应用于医学和公共卫生中非常重要的系统。