BIOPHYSICAL STUDIES ON PROTEIN FOLDING BY MASS SPECTROMETRY

近年来，质谱法已开始被用作 方法来监测蛋白质折叠的动力学。 我们将探讨 MS的这一方面使用至少两种不同的方法：（i） 通过限制性蛋白水解检测受保护的蛋白质结构域 从SDS-PAGE凝胶中分离受保护物种并鉴定 使用进一步消化和MALDI或ESI-MS。 在生理pH下的H/D交换动力学（快速交换）如下 通过胃蛋白酶消化在低pH值（缓慢交换）和鉴定 保护区作为时间的函数。 我们将调查 将这些技术应用于生物学问题，例如 鉴定由基因突变引起的不同折叠， 类固醇生成急性调节蛋白星星。

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.