Native Mass Spectrometry for Structural Biophysics
Native Mass Spectrometry for Structural Biophysics
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用于结构生物物理学的天然质谱分析
DOI：
10.1016/j.bpj.2013.11.032
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发表时间：
2014
影响因子：
3.4
通讯作者：
Benesch J
中科院分区：
文献类型：
--
作者：
Benesch J
我们使用基于质谱（MS）的方法组合，直接询问100 kDa至1 MDa范围内蛋白质的四级结构和动力学，在真空中保持完整[1]。我们特别感兴趣的是小的热休克蛋白分子伴侣，它负责确保蛋白质在细胞中到达并保持其天然折叠[2]。然而，由于其频繁的异质性和运动的平衡，他们的研究往往受到阻碍。MS和串联MS使我们能够识别和定量的相对丰度不同的蛋白质化学计量存在于溶液中，而实时实验允许提取的四元波动。这导致我们获得的基本蛋白质-蛋白质和蛋白质-配体相互作用的平衡和速率常数。同时，离子迁移率（IM）MS测量提供了有关蛋白质物理大小的信息。与其他来源的信息一起，这些实验因此提供了强有力的限制，在建模蛋白质组装体的结构，这是困难和耗时的研究，通过传统的结构生物学方法。
We use a combination of mass spectrometry (MS) based approaches to interrogate directly the quaternary structure and dynamics of proteins in the 100 kDa to 1 MDa range, intact in vacuum [1]. Of particular interest to us are the small heat-shock protein molecular chaperones, which are responsible for ensuring proteins reach and maintain their native fold in the cell [2]. Their study however is often hampered however due to their frequent heterogeneity and motions at equilibrium.MS and tandem MS enable us to identify and quantify the relative abundances of different protein stoichiometries present in solution, while real-time experiments allow the extraction of quaternary fluctuations. This leads us to obtain equilibrium and rate constants for the underlying protein-protein and protein-ligand interactions. Concurrently, ion mobility (IM) MS measurements provide information as to the physical size of the proteins. Together with information from other sources, these experiments therefore provide powerful restraints in modeling the structures of protein assemblies that are difficult and time-consuming to study by means of conventional structural biology approaches.