Purification and kinetic characterization of human indoleamine 2,3-dioxygenases 1 and 2 (IDO1 and IDO2) and discovery of selective IDO1 inhibitors
Purification and kinetic characterization of human indoleamine 2,3-dioxygenases 1 and 2 (IDO1 and IDO2) and discovery of selective IDO1 inhibitors
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DOI：
10.1016/j.bbapap.2011.07.023
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发表时间：
2011-12-01
影响因子：
3.2
通讯作者：
Sutherland, Claire L.
中科院分区：
文献类型：
--
作者：
Meininger, David;Zalameda, Leeanne;Sutherland, Claire L.
吲哚胺2，3-双加氧酶(IDO1)催化沿着犬尿氨酸途径分解色氨酸的第一步。由于IDO1在癌症、低血压和神经退行性疾病等多种疾病的发病机制中的作用，IDO1的治疗抑制受到了极大的关注。一种相关的酶，IDO2，最近已经被描述。我们首次报道了利用一种简便易行的t-色氨酸消耗试验对人IDO2进行纯化和动力学表征，该方法适用于高通量筛选。我们发现，人IDO2对t-色氨酸的Km远高于IDO1。我们还描述了一种新的IDO1抑制剂化合物AMG-1的高通量筛选的鉴定和表征，并比较了IDO1和IDO2与AMG-1和先前报道的化合物的抑制谱。我们的数据表明，人IDO1和IDO2具有不同的动力学参数和不同的抑制曲线。AMG-1和相关类似物与IDO1的已知结构和同源建模的IDO2的对接表明了IDO1和IDO2不同抑制谱的可能原理。(C)2011爱思唯尔B.V.保留所有权利。
Indoleamine 2,3-dioxygenase (IDO1) catalyzes the first step in tryptophan breakdown along the kynurenine pathway. Therapeutic inhibition of IDO1 is receiving much attention due to its proposed role in the pathogenesis of several diseases including cancer, hypotension and neurodegenerative disorders. A related enzyme, IDO2 has recently been described. We report the first purification and kinetic characterization of human IDO2 using a facile t-tryptophan consumption assay amenable to high throughput screening. We found that the Km of human IDO2 for t-tryptophan is much higher than that of IDO1. We also describe the identification and characterization of a new IDO1 inhibitor compound, Amg-1, by high throughput screening, and compare the inhibition profiles of IDO1 and IDO2 with Amg-1 and previously described compounds. Our data indicate that human IDO1 and IDO2 have different kinetic parameters and different inhibition profiles. Docking of Amg-1 and related analogs to the known structure of IDO1 and to homology-modeled IDO2 suggests possible rationales for the different inhibition profiles of IDO1 and IDO2. (C) 2011 Elsevier B.V. All rights reserved.