Dynamics of protein kinases and pseudokinases by HDX-MS.
Dynamics of protein kinases and pseudokinases by HDX-MS.
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DOI：
10.1016/bs.mie.2022.03.037
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发表时间：
2022
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蛋白激酶折叠的动态与其结构密切相关。过去三十年的激酶生物物理研究揭示了激酶结构域的关键动态特征，以及最近这些特征如何赋予催化受损的激酶-或假激酶-信号特性。氢-氚交换-质谱联用(HDX-MS)被证明是一种有价值的方法，可以用来详细研究激酶和假激酶的动力学。在这里，我们简要讨论了提供对蛋白激酶动力学的洞察的方法，描述了HDX-MS是如何被用来回答在激酶/假激酶领域中的问题的，并提供了收集HDX-MS数据集以研究小分子结合到假激活域的影响的详细协议。随着更多的假酶构象干扰小分子的发现，HDX-MS很可能成为探索药物诱导的假酶动力学变化的一种强有力的方法。
Dynamics of the protein kinase fold are deeply intertwined with its structure. The past three decades of kinase biophysical studies revealed the key dynamic features of the kinase domain and, more recently, how these features may endow catalytically impaired kinases—or pseudokinases—with signaling properties. Hydrogen-deuterium exchange coupled with mass spectrometry (HDX-MS) is proving to be a valuable approach for detailed studies of kinase and pseudokinase dynamics. Here we briefly discuss the methods that have provided insights into protein kinase dynamics, describe how HDX-MS is being used to answer questions in the kinase/pseudokinase field, and provide a detailed protocol for collecting an HDX-MS dataset to study the impacts of small molecule binding to a pseudokinase domain. As more pseudokinase conformational disrupting small molecules are discovered, HDX-MS is likely to be a powerful approach for exploring drug-induced changes in pseudokinase dynamics.