Inhibition of thioredoxin reductase 1 by porphyrins and other small molecules identified by a high-throughput screening assay.
Inhibition of thioredoxin reductase 1 by porphyrins and other small molecules identified by a high-throughput screening assay.
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DOI：
10.1016/j.freeradbiomed.2011.01.020
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发表时间：
2011-05-01
影响因子：
7.4
通讯作者：
Simeonov, Anton
中科院分区：
文献类型：
--
作者：
Prast-Nielsen, Stefanie;Dexheimer, Thomas S.;Schultz, Lena;Stafford, William C.;Cheng, Qing;Xu, Jianqiang;Jadhav, Ajit;Arner, Elias S. J.;Simeonov, Anton
关键词：
硒蛋白硫氧还蛋白还原酶1（TrxR 1）是近年来发现的一个很有前途的抗癌药物靶点。因此，需要高通量测定来发现靶向酶的新型化合物。在此，我们描述了一种单酶，双目的测定同时识别抑制剂和底物的TrxR 1。使用该试验筛选LOPAC 1280化合物集合，我们鉴定了几种已知的TrxR 1抑制剂，从而验证了该试验，以及几种迄今未知的靶向酶的化合物。这些包括rottlerin（以前报道为PKC δ抑制剂和线粒体解偶联剂）和血红素前体原卟啉IX（PpIX）。我们发现PpIX是TrxR 1的一个强有力的竞争性抑制剂，相对于Trx 1的Ki = 2.7 μM，在没有Trx 1的情况下，表现出时间依赖性的不可逆抑制，表观二级速率常数（kinact）为0.73×10−3 ± 0.07×10−3 μM− 1 min −1。外源递送的PpIX具有细胞毒性，抑制A549细胞增殖，并且发现还抑制细胞TrxR活性。氯化血红素和亚铁螯合酶抑制剂NMPP也抑制TrxR 1，并表现出细胞毒性，但较PpIX效力低。我们得出结论，罗特勒诱导的细胞效应可能涉及靶向TrxR 1。PpIX作为TrxR 1抑制剂的意外发现表明，这种抑制可能有助于与异常高PpIX水平的病症相关的症状;例如，在红细胞生成性原卟啉症中观察到的亚铁螯合酶活性降低。最后，基于本文提出的新的高通量TrxR 1测定，可以发现和进一步表征TrxR 1的其他抑制剂。
The selenoprotein thioredoxin reductase 1 (TrxR1) has in recent years been identified as a promising anticancer drug target. A high throughput assay for discovery of novel compounds targeting the enzyme is therefore warranted. Herein, we describe a single-enzyme, dual-purpose assay for simultaneous identification of inhibitors and substrates of TrxR1. Using this assay to screen the LOPAC1280 compound collection we identified several known inhibitors of TrxR1, thus validating the assay, as well as several compounds hitherto unknown to target the enzyme. These included rottlerin (previously reported as a PKC delta inhibitor and mitochondrial uncoupler) and the heme precursor protoporphyrin IX (PpIX). We found that PpIX was a potent competitive inhibitor of TrxR1 with a Ki = 2.7 μM with regards to Trx1, and in the absence of Trx1 displayed time dependent irreversible inhibition with an apparent second-order rate constant (kinact) of 0.73×10−3 ± 0.07×10−3 μM−1min−1. Exogenously delivered PpIX was cytotoxic, inhibited A549 cell proliferation and was found to also inhibit cellular TrxR activity. Hemin and the ferrochelatase inhibitor NMPP also inhibited TrxR1 and showed cytotoxicity, but less potently compared to PpIX. We conclude that rottlerin-induced cellular effects may involve targeting of TrxR1. The unexpected finding of PpIX as a TrxR1 inhibitor suggests that such inhibition may contribute to symptoms associated with conditions of abnormally high PpIX levels; such as, reduced ferrochelatase activity seen in erythropoietic protoporphyria. Finally, additional inhibitors of TrxR1 may be discovered and further characterized based upon the new high throughput TrxR1 assay presented here.
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影响因子：
4.8
作者：
Cenas, N;Prast, S;Arnér, ESJ
通讯作者：
Arnér, ESJ
DOI：
10.2174/1568011043352984
发表时间：
2004-05-01
期刊：
Current Medicinal Chemistry - Anti-Cancer Agents
影响因子：
--
作者：
Collaud, Sabine;Juzeniene, Asta;Lange, Norbert
通讯作者：
Lange, Norbert
影响因子：
4.8
作者：
Gromer, S;Arscott, LD;Becker, K
通讯作者：
Becker, K
影响因子：
7.4
作者：
Arnér, ESJ;Nakamura, H;Spyrou, G
通讯作者：
Spyrou, G
DOI：
10.1161/01.atv.11.6.1700
发表时间：
1991-11-01
期刊：
ARTERIOSCLEROSIS AND THROMBOSIS
影响因子：
--
作者：
BALLA, G;JACOB, HS;VERCELLOTTI, GM
通讯作者：
VERCELLOTTI, GM