QUANTITATION OF OXIDATION-INDUCED CONFORMATIONAL CHANGES BY 2ND ORDER REACTIONS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 最近，一种利用蛋白质的闪光快速光氧化(FPOP)来研究羟基足迹的方法被发表，该方法声称完成氧化化学的速度快于蛋白质能够经历显著的氧化诱导的去折叠。不幸的是，验证这一说法的方法本质上是间接的，方法是将氧化部位与已公布的高分辨结构进行比较。为了验证FPOP方法可以比蛋白质展开更快地完成氧化化学，我们正在开发一种方法，通过分析完整的蛋白质氧化产物来检测和定量二级羟基自由基足迹实验中发生的氧化诱导展开的量。

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. Recently, a method for hydroxyl radical footprinting using flash fast photooxidation of proteins (FPOP) was published that claims to complete the oxidation chemistry faster than the protein can undergo a significant oxidation-induced unfolding. Unfortunately, the methods for verifying this claim were indirect in nature, by comparing sites of oxidation with published high resolution structures. In order to verify that the FPOP method can complete oxidation chemistry faster than the protein can unfold, we are developing a method for detecting and quantitating the amount of oxidation-induced unfolding that occurs during a second order hydroxyl radical footprinting experiment using analysis of the intact protein oxidation products.