Covalent Modifier Discovery Using Hydrogen/Deuterium Exchange-Mass Spectrometry
Covalent Modifier Discovery Using Hydrogen/Deuterium Exchange-Mass Spectrometry
复制标题
使用氢/氘交换质谱法发现共价修饰剂
DOI：
10.1021/acs.jmedchem.2c01986
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发表时间：
2023
影响因子：
7.3
通讯作者：
Itoh Toshimasa
中科院分区：
文献类型：
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作者：
Kojima Hiroyuki;Yanagi Ryota;Higuchi Eri;Yoshizawa Mami;Shimodaira Tomoyuki;Kumagai Misaki;Kyoya Tatsuhiro;Sekine Miyu;Egawa Daichi;Ohashi Nami;Ishida Hiroaki;Yamamoto Keiko;Itoh Toshimasa
共价配体通常从用于高通量筛选的化学库中过滤出，因为亲电官能团被认为是泛测定干扰化合物（PAINS）。因此，筛选策略，可以区分真正的共价配体从PAINS是必要的。氢/氘交换质谱（HDX-MS）是一种评价蛋白质稳定性的有力工具。在这里，我们报告了一个共价修饰剂的筛选方法，使用HDX-MS。在这项研究中，HDX-MS用于分类过氧化物酶体增殖物激活受体γ（PPARγ）和维生素D受体配体。HDX-MS可以区分配体-蛋白质相互作用的强度。我们的HDX-MS筛选方法鉴定了LT 175和nTZDpa，它们可以同时结合到具有协同激活作用的PPARγ配体结合结构域（PPARγ-LBD）。此外，碘乙酸被鉴定为稳定PPARγ-LBD的新型共价修饰剂。
Covalent ligands are generally filtered out of chemical libraries used for high-throughput screening, because electrophilic functional groups are considered to be pan-assay interference compounds (PAINS). Therefore, screening strategies that can distinguish true covalent ligands from PAINS are required. Hydrogen/deuterium–exchange mass spectrometry (HDX–MS) is a powerful tool for evaluating protein stability. Here, we report a covalent modifier screening approach using HDX–MS. In this study, HDX–MS was used to classify peroxisome proliferator-activated receptor γ (PPARγ) and vitamin D receptor ligands. HDX–MS could discriminate the strength of ligand–protein interactions. Our HDX–MS screening method identified LT175 and nTZDpa, which can bind concurrently to the PPARγ ligand-binding domain (PPARγ–LBD) with synergistic activation. Furthermore, iodoacetic acid was identified as a novel covalent modifier that stabilizes the PPARγ–LBD.