Ling Jessmi M L, Silva Leslie, Schriemer David C, Schryvers Anthony B
Department of Microbiology and Infectious Diseases, University of Calgary, Calgary, AB, Canada.
Methods Mol Biol. 2012;799:237-52. doi: 10.1007/978-1-61779-346-2_15.
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.
描述了一种利用氢/氘交换与质谱联用探索蛋白质-蛋白质相互作用的方法。该方法监测重水中溶液中主链（酰胺）氢的交换，这种交换主要发生在蛋白质暴露于溶剂的部分。在存在蛋白质结合伴侣的情况下，交换减少的区域要么是蛋白质-蛋白质相互作用界面的一部分，要么经历构象变化以减少对溶剂的可及性。该方法的优点是可在生理条件下使用未修饰的蛋白质。在本章中，我们描述了一种适用于使用传统质谱系统探测相对较大蛋白质之间相互作用的方法。以人转铁蛋白与脑膜炎奈瑟菌受体蛋白转铁蛋白结合蛋白B之间的相互作用为例，提供了一个具有挑战性的系统。
