Electrophoretic Measurements of Protein Folding Intermediates

德克萨斯大学奥斯汀分校的Jason Shear教授将与Eric Anslyn教授和Andrew Ellington教授合作开展一个由分析和表面化学项目支持的生物分析化学项目。PI在他之前的职业计划中发起了门控超快速电泳分离方法。这个想法是把一个分子笼起来，光化学激活它，在微秒内进行分离，然后用光来检测化合物，就像飞行时间一样。这个项目的重点是将这种方法应用于蛋白质折叠问题。新的光化学触发与帮助位点定向诱变，以实现所需的化学正在探索。将使用两种模型蛋白：细胞色素c和无肌红蛋白。该项目将分析化学和物理化学、有机合成和分子生物学相结合，开发和评估一种快速灵敏检测蛋白质折叠过程的新型仪器。这项研究的动机是蛋白质序列如何指定构象的问题，这是生物学中一个悬而未决的主要问题。因此，对蛋白质再折叠的能量学和动力学的定量信息有很大的需求。错误折叠蛋白的作用与阿尔茨海默病等越来越多的疾病有关，但尚未得到很好的理解。

Professor Jason Shear of the University of Texas Austin will work with Professors Eric Anslyn and Andrew Ellington on a project in bioanalytical chemistry supported by the Analytical and Surface Chemistry Program. The PI initiated a gated ultra-fast electrophoretic separations method in his previous award in the CAREER program. The idea is to cage a molecule, activate it photochemically, do a separation in microseconds and then detect the compounds using light in a time-of-flight analogy. This project centers on the application of that method to the problem of protein folding. Novel photochemical triggering with the help of site-directed mutagenesis to achieve the desired chemistry are being explored. Two model proteins will be used: cytochrome c and apomyoglobin. The project combines analytical and physical chemistry, organic synthesis and molecular biology to develop and evaluate a novel instrument for fast and sensitive detection of protein folding processes. The research is motivated by the question of how protein sequence specifies conformation, a major unanswered question in biology. There is thus a great need for quantitative information about the energetics and dynamics of protein refolding. The role of misfolded proteins is implicated but not well understood in a growing number of diseases such as Alzheimer's disease.