Hydrogen-Deuterium Exchange Coupled to Mass Spectrometry to Investigate Ligand-Receptor Interactions
Hydrogen-Deuterium Exchange Coupled to Mass Spectrometry to Investigate Ligand-Receptor Interactions
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DOI：
10.1007/978-1-61779-346-2_15
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发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Schryvers, Anthony B.
中科院分区：
文献类型：
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作者：
Ling, Jessmi M. L.;Silva, Leslie;Schryvers, Anthony B.
本发明描述了一种使用氢/氘交换耦合到质谱来探索蛋白质蛋白质相互作用的方法。该方法监测氘代水溶液中主要发生在暴露于溶剂的蛋白质部分上的骨架（酰胺）氢的交换。在蛋白质结合配偶体的存在下，经历减少的交换的区域是蛋白质-蛋白质相互作用界面的一部分或经历构象变化以减少对溶剂的可及性。该方法具有在生理条件下使用未经修饰的蛋白质的优点。在这一章中，我们描述了一种适合探测相对较大的蛋白质之间的相互作用，使用传统的质谱系统的方法。人转铁蛋白和脑膜炎奈瑟氏球菌受体蛋白、转铁蛋白结合蛋白B之间的相互作用提供了一个具有挑战性的系统作为例子。
A method for exploring protein protein interactions using hydrogen/deuterium exchange coupled to mass spectrometry is described. The method monitors the exchange of backbone (amide) hydrogens in solutions of deuterated water that primarily occur on portions of the protein exposed to solvent. In the presence of a protein binding partner, regions that experience reduced exchange are either part of the protein protein interaction interface or undergo conformational changes to reduce accessibility to solvent. This method has the advantage of being used under physiological conditions with unmodified proteins. In this chapter, we describe an approach suitable for probing interactions among relatively large proteins using conventional mass spectrometry systems. The interaction between human transferrin and the Neisseria meningitidis receptor protein, transferrin binding protein B, provides a challenging system as an example.