Synergistic Structural Information from Covalent Labeling and Hydrogen-Deuterium Exchange Mass Spectrometry for Protein Ligand Interactions
Synergistic Structural Information from Covalent Labeling and Hydrogen-Deuterium Exchange Mass Spectrometry for Protein Ligand Interactions
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DOI：
10.1021/acs.analchem.9b04257
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发表时间：
2019-12-03
影响因子：
7.4
通讯作者：
Vachet, Richard W.
中科院分区：
文献类型：
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作者：
Liu, Tianying;Limpikirati, Patanachai;Vachet, Richard W.
氢氘交换（HDX）质谱（MS）和共价标记（CL）MS通常被认为是用于蛋白质结构分析的互补方法，因为一种方法探测蛋白质骨架，而另一种方法探测侧链。对于蛋白质配体的相互作用，我们证明在这项工作中，这两种标记技术可以提供协同的结构信息，蛋白质配体结合时，如焦碳酸二乙酯（DEPC）的试剂用于CL，因为DEPC和HDX的反应速率的差异。使用三个模型蛋白质配体系统，我们表明，DEPC标记的时间尺度较慢，使其只敏感于溶剂的可及性和蛋白质结构波动的变化不敏感的变化，而HDX是敏感的溶剂的可及性和结构波动的变化。当一起使用时，这两种方法更清楚地揭示结合位点和配体诱导的远离结合位点的结构波动变化，这比单独使用任何一种技术都能提供更全面的信息。我们预测，这两种方法将发现广泛的使用更深入地了解蛋白质配体相互作用。
Hydrogen deuterium exchange (HDX) mass spectrometry (MS) and covalent labeling (CL) MS are typically considered to be complementary methods for protein structural analysis, because one probes the protein backbone, while the other probes side chains. For protein ligand interactions, we demonstrate in this work that the two labeling techniques can provide synergistic structural information about protein ligand binding when reagents like diethylpyrocarbonate (DEPC) are used for CL because of the differences in the reaction rates of DEPC and HDX. Using three model protein ligand systems, we show that the slower time scale for DEPC labeling makes it only sensitive to changes in solvent accessibility and insensitive to changes in protein structural fluctuations, whereas HDX is sensitive to changes in both solvent accessibility and structural fluctuations. When used together, the two methods more clearly reveal binding sites and ligand-induced changes to structural fluctuations that are distant from the binding site, which is more comprehensive information than either technique alone can provide. We predict that these two methods will find widespread usage together for more deeply understanding protein ligand interactions.