Protein dynamics studies by hydrogen exchange

描述(申请人提供)：在某些情况下，氢交换测量已经能够区分不同种类的蛋白质运动，并提供关于它们的特性、它们的程度、它们的平衡和动力学参数、它们对环境条件和突变的反应以及它们与功能特性的关系的现场分辨信息。作为理解蛋白质结构动态维度的意义及其在蛋白质功能中的作用的努力的一部分，这些氢交换能力需要改进并扩展到更多的蛋白质。最初建议研究葡萄球菌核酸酶、二氢叶酸还原酶和细胞色素c，以及更广泛的其他蛋白质。为了更好地了解将氢交换行为与蛋白质结构和动力学联系起来的关系，将使用大量结果来测试和改进试图将蛋白质性质和氢交换联系起来的理论模型。为了扩大天然态氢交换方法的范围，渗透分子的作用模式及其作为逆变性剂的用途将被探索。此外，还将研究更大的蛋白质，这些蛋白质有望改变局部波动和通过氢交换测量的更大展开反应之间的平衡。为了研究蛋白质结构的动态维度的功能作用，将氢交换方法应用于酶功能的研究。

DESCRIPTION (provided by applicant): Hydrogen exchange measurements have been able, in some cases, to distinguish different kinds of protein motions and to provide site-resolved information on their identity, their extent, their equilibrium and kinetic parameters, their response to ambient conditions and mutations, and their relationship to functional properties. As part of the effort to understand the significance of the dynamic dimension of protein structure and its role in protein function, these hydrogen exchange capabilities need to be improved and extended to more proteins. Studies are proposed of staphylococcal nuclease, dihydrofolate reductase, and cytochrome c initially, and other proteins more broadly. To better understand the relationships that connect hydrogen exchange behavior to protein structure and dynamics, a large quantity of results will be used to test and refine theoretical models that attempt to relate protein properties and hydrogen exchange. To extend the scope of native state hydrogen exchange methods, the mode of action of osmolytes, and their use as reverse denaturants, will be pursued. Also, larger proteins, which promise to change the balance between local fluctuations and larger unfolding reactions measured by hydrogen exchange, will be studied. To study the functional role of the dynamic dimension of protein structure, hydrogen exchange approaches will be applied to the study of enzymatic function.