BIOPHYSICAL STUDIES ON PROTEIN FOLDING BY MASS SPECTROMETRY

近年来，质谱学已开始被用作 方法监测蛋白质折叠的动态变化。我们将探索 MS的这一方面使用至少两种不同的方法：(I) 有限蛋白水解法检测受保护的蛋白质结构域 从SDS-PAGE凝胶中分离和鉴定受保护物种 使用进一步消化和MALDI或ESI-MS。(Ii)监察 在生理pH(快速交换)下H/D交换的动力学 低pH(慢交换)胃酶消化法及其鉴定 受保护区域作为时间的函数。我们将调查 这些技术在生物学问题上的应用，例如 基因突变引起的不同褶皱的鉴定 类固醇合成急性调节蛋白STAR。

In recent years mass spectrometry has started to be used as a method to monitor the dynamics of protein folding. We will explore this aspect of MS using at least two different approaches: (i) The detection of protected protein domains by limited proteolytic isolation of protected species from SDS-PAGE gels and identification using further digestion and MALDI or ESI-MS. (ii) monitoring the kinetics of H/D exchange at physiological pH (fast exchange) followed by pepsin digestion at low pH (slow exchange) and identification of protected regions as a function of time. We will investigate the application of these techniques to biological problems, such as identification of different folds caused by mutations in the steroidogenic acute regulatory protein StAR.