DMSO-perturbing assay for identifying promiscuous enzyme inhibitors
DMSO-perturbing assay for identifying promiscuous enzyme inhibitors
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用于鉴定混杂酶抑制剂的 DMSO 扰动测定
DOI：
10.1021/acsmedchemlett.9b00093
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发表时间：
2019
影响因子：
4.2
通讯作者：
Takeru Nose.
中科院分区：
文献类型：
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作者：
Keisuke Tomohara;Isao Adachi;Yoshikazu Horino;Hitoshi Kesamaru;Hitoshi Abe;Keitaro Suyama;Takeru Nose.
在寻找酶抑制剂的过程中，我们经常会遇到“混杂”的酶抑制剂，它们表现出与酶活性部位的非特异性结合性质。因此，在发现过程中，应尽早对抑制性候选基因进行机械性表征。然而，仍然缺乏高度可靠和容易获得的方法来评估初始HITS抑制剂的特异性。本研究发展并建立了一种新的DMSO干扰试验来鉴定混杂酶抑制剂。该方法成功地鉴定了范围广泛的非特异性结合抑制剂，通常是通过DMSO加成影响抑制活性的减弱来实现的。这种衰减可以归因于抑制剂对扰动溶液中酶的产生态和非产生态(非变性)的非特异性结合特性。这一工作假说得到了酶构象的光谱分析和溶剂对扰动影响的分析的支持。总体而言，这些结果为DMSO干扰试验提供了一个新的概念。
In search for enzyme inhibitors, we often encounter “promiscuous” enzyme inhibitors exhibiting nonspecific binding property toward enzyme active site. Therefore, inhibitory candidates should be mechanistically characterized as early as possible in discovery processes. However, there remains a lack of highly reliable and readily available methodology to evaluate specificity of initial hits inhibitors. The present study developed and established a novel DMSO-perturbing assay to identify promiscuous enzyme inhibitors. The assay successfully identified nonspecific binding inhibitors with a broad scope, typically by the attenuation of inhibitory activity by the influence of DMSO-addition. This attenuation would be attributed to the nonspecific binding property of inhibitors toward both productive and nonproductive (nondenatured) states of enzymes in perturbation solution. This working hypothesis was supported by spectroscopic analyses of enzyme conformations and analyses of solvent effects on perturbation. Overall, these results provided a novel concept of the DMSO-perturbing assay.