Native mass spectrometry: towards high-throughput structural proteomics.
Native mass spectrometry: towards high-throughput structural proteomics.
复制标题
DOI：
10.1007/978-1-4939-2230-7_18
复制
发表时间：
2015-01-01
期刊：
影响因子：
--
通讯作者：
Benesch, Justin L P
中科院分区：
文献类型：
--
作者：
Kondrat, Frances D L;Struwe, Weston B;Benesch, Justin L P
天然质谱 (MS) 已成为结构蛋白质组学的一种敏感方法，使从业者能够深入了解蛋白质自组装，包括化学计量和三维结构，以及互补的热力学和动力学方面。尽管质谱通常在真空中进行，但大量文献描述了如何在实验的时间尺度上很大程度上保留天然溶液态结构。天然 MS 的优点是，与 NMR 和 X 射线晶体学等传统结构技术相比，它需要的样品数量少得多，因此非常适合高通量研究。在这里，我们首先描述原生 MS 方法并概述它可以提供的结构蛋白质组数据。然后，我们提供实验的实用细节，以检查蛋白质组装体的结构和动态特性，强调潜在的陷阱以及最佳实践的原则。
Native mass spectrometry (MS) has become a sensitive method for structural proteomics, allowing practitioners to gain insight into protein self-assembly, including stoichiometry and three-dimensional architecture, as well as complementary thermodynamic and kinetic aspects. Although MS is typically performed in vacuum, a body of literature has described how native solution-state structure is largely retained on the timescale of the experiment. Native MS offers the benefit that it requires substantially smaller quantities of a sample than traditional structural techniques such as NMR and X-ray crystallography, and is therefore well suited to high-throughput studies. Here we first describe the native MS approach and outline the structural proteomic data that it can deliver. We then provide practical details of experiments to examine the structural and dynamic properties of protein assemblies, highlighting potential pitfalls as well as principles of best practice.