RADICAL FOOTPRINTING BY SUB-MICROSECOND ELECTRON BEAM PULSE

亚微秒电子束脉冲的自由基足迹

• 批准号: 8361800
• 负责人: JOSHUA S SHARP
• 金额: $ 3.54万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361800
• 关键词: Carbohydrates; Complex; Electron Beam; Electrons; Event; Funding; Grant; Hydrogen Peroxide; Hydroxyl Radical; Lasers; Metals; Methods; National Center for Research Resources; Organized by Structure Protein; Oxidants; Physiologic pulse; Principal Investigator; Proteins; Protocols documentation; Reaction; Research; Research Infrastructure; Resources; Solvents; Source; Techniques; United States National Institutes of Health; Water; Work; cost; oxidation; photolysis; 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. Hydroxyl radical footprinting is a new method for studying protein structure by examining differences in the rate of hydroxyl radical-protein sidechain reaction between a free protein and a protein-carbohydrate complex, and correlating changes in the rate of oxidation with changes in solvent accessibility. In order to probe only the native structure of the protein-carbohydrate complex, one must either strictly limit the reaction to one oxidation event per protein-carbohydrate complex, or limit the timescale of the reaction to complete faster than large scale protein conformational changes can occur. It has previously been shown that laser-induced photolysis of hydrogen peroxide can result in heavy oxidation that can be controlled to be completed on the sub-microsecond timescale. Unfortunately, this protocol requires high concentrations of hydrogen peroxide, which is capable of directly oxidizing proteins by both metal-catalyzed and metal-free mechanisms, on very fast timescales. We are working to develop a technique to oxidize proteins on a sub-microsecond timescale without the use of a precursor oxidant by radiolysis of water using an electron pulse from a Van de Graaff accelerator.

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。 羟基自由基足迹是一种通过检查游离蛋白质与蛋白质 - 碳水化合物复合物之间的羟基自由基蛋白侧chain反应的差异来研究蛋白质结构的新方法，并将氧化速率与溶剂可及性的变化相关。 为了仅探测蛋白质 - 碳水化合物复合物的天然结构，必须严格限制每个蛋白质 - 碳水化合物复合物对一个氧化事件的反应，或者限制反应的时间尺度以比大规模蛋白质构象变化更快地完成。 以前已经显示，激光诱导的过氧化氢的光解会导致重氧化，可以控制在亚微秒次时尺度上完成。 不幸的是，该方案需要高浓度的过氧化氢，在非常快的时间标准上，能够通过金属催化和无金属机制直接氧化蛋白质。 我们正在努力开发一种技术，以在亚微秒时间尺度上氧化蛋白质，而无需通过使用Van de Graaff加速器的电子脉冲对水进行放射透明剂，而无需使用前体氧化剂。