Probing specific ligand-protein interactions by native-denatured exchange mass spectrometry
Probing specific ligand-protein interactions by native-denatured exchange mass spectrometry
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通过天然变性交换质谱法探测特定配体-蛋白质相互作用
DOI：
10.1016/j.aca.2018.07.072
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发表时间：
2018
影响因子：
6.2
通讯作者：
Ye Hui
中科院分区：
文献类型：
--
作者：
Zheng Qiuling;Tian Yang;Ruan Xujun;Chen Hao;Wu Xunxun;Xu Xiaowei;Wang Guangji;Hao Haiping;Ye Hui
探测配体-靶蛋白相互作用为深入理解生物化学机制和设计药物筛选试验提供了重要信息。天然电喷雾电离质谱（ESI-MS）是有前途的配体-蛋白质复合物的直接分析。然而，它缺乏区分特异性和非特异性配体-蛋白质相互作用的能力，以及进一步识别特异性结合的蛋白质作为药物靶点候选物的能力，这仍然是迄今为止药物开发领域的主要挑战。本文报道了一种采用液相样品解吸电喷雾电离（LS-DESI）的天然变性交换（NDX）质谱（MS）采集方法，并证明了其能够从非共价配体-蛋白质复合物的天然检测转变为使用三种模型配体-蛋白质复合物（包括肌红蛋白、CDP-核糖核酸酶和N，N′，N″-三乙酰壳三糖（NAG 3）-溶菌酶）的变性分析。值得注意的是，我们发现NDX-MS方法可以很容易地区分特异性配体-蛋白质相互作用与非特异性配体-蛋白质相互作用，如其不同的动态曲线所揭示的那样，Kdas是DESI喷雾流速的函数。因此，这种NDX-MS方法有望在未来应用于发现感兴趣配体的特异性蛋白质靶点，以及筛选对药物靶点具有高特异性的化合物，从而消除脱靶效应。
Probing ligand-target protein interactions provides essential information for deep understanding of biochemical machinery and design of drug screening assays. Native electrospray ionization-mass spectrometry (ESI-MS) is promising for direct analysis of ligand-protein complexes. However, it lacks the ability to distinguish between specific and non-specific ligand-protein interactions, and to further recognize the specifically bound proteins as drug target candidates, which remains as a major challenge in the field of drug developments by far. Herein we report a native-denatured exchange (NDX) mass spectrometry (MS) acquisition approach using a liquid sample-desorption electrospray ionization (LS-DESI) setup, and demonstrate its capability in enabling a change from native detection of noncovalent ligand-protein complexes to denatured analysis using three model ligand-protein complexes including myoglobin, CDP-ribonuclease and N,N′,N″-triacetylchitotriose (NAG3)-lysozyme. Notably, we found the NDX-MS approach can readily discriminate specific ligand-protein interactions from nonspecific ones, as revealed by their distinct dynamic profiles of Kdas a function of the DESI spraying flow rate. Consequently, this NDX-MS approach holds promise for future applications to discovering specific protein targets for ligands of interest, and to screening compounds with high specificity to drug targets and thus eliminates off-target effects.