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Examination of SOD1 aggregation modulators and their effect on SOD1 enzymatic activity as a proxy for potential toxicity
The FASEB Journal ( IF 4.4 ) Pub Date : 2020-07-23 , DOI: 10.1096/fj.202000948r
Ravinder Malik 1 , Christian Corrales 1 , Miriam Linsenmeier 1 , Huda Alalami 1 , Niki Sepanj 1 , Gal Bitan 1, 2, 3
Affiliation  

Small‐molecule inhibitors of abnormal protein self‐assembly are promising candidates for developing therapy against proteinopathies. Such compounds have been examined primarily as inhibitors of amyloid β‐protein (Aβ), whereas testing of inhibitors of other amyloidogenic proteins has lagged behind. An important issue with screening compound libraries is that although an inhibitor suitable for therapy must be both effective and nontoxic, typical screening focuses on efficacy, whereas safety typically is tested at a later stage using cells and/or animals. In addition, typical thioflavin T (ThT)‐fluorescence‐based screens use the final fluorescence value as a readout, potentially missing important kinetic information. Here, we examined potential inhibitors of superoxide dismutase 1 (SOD1) using ThT‐fluorescence including the different phases of fluorescence change and added a parallel screen of SOD1 activity as a potential proxy for compound toxicity. Some compounds previously reported to inhibit other amyloidogenic proteins also inhibited SOD1 aggregation at low micromolar concentrations, whereas others were ineffective. Analysis of the lag phase and exponential slope added important information that could help exclude false‐positive or false‐negative results. SOD1 was highly resistant to inhibition of its activity, and therefore, did not have the necessary sensitivity to serve as a proxy for examining potential toxicity.

中文翻译:

检查 SOD1 聚集调节剂及其对 SOD1 酶活性的影响作为潜在毒性的代表

异常蛋白质自组装的小分子抑制剂是开发针对蛋白质病的疗法的有希望的候选者。此类化合物主要作为淀粉样蛋白 β 蛋白 (Aβ) 的抑制剂进行检查,而对其他淀粉样蛋白抑制剂的测试则滞后。筛选化合物库的一个重要问题是,尽管适用于治疗的抑制剂必须既有效又无毒,但典型的筛选侧重于功效,而安全性通常在后期使用细胞和/或动物进行测试。此外,典型的基于硫代黄素 T (ThT) 荧光的屏幕使用最终荧光值作为读数,可能会丢失重要的动力学信息。这里,我们使用 ThT 荧光检查了潜在的超氧化物歧化酶 1 (SOD1) 抑制剂,包括荧光变化的不同阶段,并添加了 SOD1 活性的平行筛选作为化合物毒性的潜在代表。一些先前报道的抑制其他淀粉样蛋白的化合物也在低微摩尔浓度下抑制 SOD1 聚集,而其他化合物则无效。滞后阶段和指数斜率的分析增加了有助于排除假阳性或假阴性结果的重要信息。SOD1 对其活性的抑制具有高度抗性,因此,它没有必要的敏感性来作为检测潜在毒性的代理。一些先前报道的抑制其他淀粉样蛋白的化合物也在低微摩尔浓度下抑制 SOD1 聚集,而其他化合物则无效。滞后阶段和指数斜率的分析增加了有助于排除假阳性或假阴性结果的重要信息。SOD1 对其活性的抑制具有高度抗性,因此,它没有必要的敏感性来作为检测潜在毒性的代理。一些先前报道的抑制其他淀粉样蛋白的化合物也在低微摩尔浓度下抑制 SOD1 聚集,而其他化合物则无效。滞后阶段和指数斜率的分析增加了有助于排除假阳性或假阴性结果的重要信息。SOD1 对其活性的抑制具有高度抗性,因此,它没有必要的敏感性来作为检测潜在毒性的代理。
更新日期:2020-07-23
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