Many proteins do not exist in a single rigid conformation. Protein motions, or dynamics, exist and in many cases are important for protein function. The analysis of protein dynamics relies on biophysical techniques that can distinguish simultaneously existing populations of molecules and their rates of interconversion. Hydrogen exchange (HX) detected by mass spectrometry (MS) is contributing to our understanding of protein motions by revealing unfolding and dynamics on a wide timescale, ranging from seconds to hours to days. In this review, we discuss HX MS-based analyses of protein dynamics, using our studies of multi-domain kinases as examples. Using HX MS, we have successfully probed protein dynamics and unfolding in the isolated SH3, SH2 and kinase domains of the c-Src and Abl kinase families, as well as the role of intermolecular and intramolecular interactions in the global control of kinase function. Coupled with high-resolution structural information, HX MS has proved to be a powerful and versatile tool for the analysis of the conformational dynamics in these kinase systems, and has provided fresh insight regarding the regulatory control of these important signalling proteins. HX MS studies of dynamics are applicable not only to the proteins we illustrate here, but to a very wide range of proteins and protein systems, and should play a role in both the classification of and greater understanding of the prevalence of protein motion.

中文翻译：

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通过氢交换质谱法观察到的蛋白质中的部分协同展开

许多蛋白质不存在于单一的刚性构象中。存在蛋白质运动或动力学，并且在许多情况下对蛋白质功能很重要。蛋白质动力学分析依赖于生物物理技术，这些技术可以同时区分现有的分子群及其相互转化率。质谱 (MS) 检测到的氢交换 (HX) 通过揭示从几秒钟到几小时到几天不等的广泛时间尺度上的展开和动力学，有助于我们理解蛋白质运动。在这篇综述中，我们以我们对多域激酶的研究为例，讨论了基于 HX MS 的蛋白质动力学分析。使用 HX MS，我们成功地探测了 c-Src 和 Abl 激酶家族的分离的 SH3、SH2 和激酶结构域中的蛋白质动力学和解折叠，以及分子间和分子内相互作用在激酶功能的全局控制中的作用。结合高分辨率结构信息，HX MS 已被证明是分析这些激酶系统中构象动力学的强大而通用的工具，并为这些重要信号蛋白的调控提供了新的见解。HX MS 动力学研究不仅适用于我们在这里说明的蛋白质，而且适用于非常广泛的蛋白质和蛋白质系统，并且应该在蛋白质运动的分类和更深入的理解中发挥作用。HX MS 已被证明是用于分析这些激酶系统中的构象动力学的强大而通用的工具，并为这些重要信号蛋白的调节控制提供了新的见解。HX MS 动力学研究不仅适用于我们在这里说明的蛋白质，而且适用于非常广泛的蛋白质和蛋白质系统，并且应该在蛋白质运动的分类和更深入的理解中发挥作用。HX MS 已被证明是用于分析这些激酶系统中的构象动力学的强大而通用的工具，并为这些重要信号蛋白的调节控制提供了新的见解。HX MS 动力学研究不仅适用于我们在这里说明的蛋白质，而且适用于非常广泛的蛋白质和蛋白质系统，并且应该在蛋白质运动的分类和更深入的理解中发挥作用。