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

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 研究中心，而研究中心不一定是研究者所在的机构。 最近，发表了一种使用蛋白质的快速光氧化（FPOP）的羟基自由基足迹法，该方法声称比蛋白质可以经历显著的氧化诱导的解折叠更快地完成氧化化学。 不幸的是，验证这一说法的方法本质上是间接的，通过比较氧化位点与已发表的高分辨率结构。 为了验证FPOP方法可以比蛋白质展开更快地完成氧化化学，我们正在开发一种用于检测和定量氧化诱导的展开量的方法，该氧化诱导的展开量在使用完整蛋白质氧化产物的分析的二阶羟基自由基足迹实验期间发生。

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.