Small molecule inhibitors of the RNA-dependent protein kinase
Small molecule inhibitors of the RNA-dependent protein kinase
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DOI：
10.1016/s0006-291x(03)01318-4
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发表时间：
2003-08-15
影响因子：
3.1
通讯作者：
Beal, PA
中科院分区：
文献类型：
--
作者：
Jammi, NV;Whitby, LR;Beal, PA
RNA依赖的蛋白激酶(PKR)是一种干扰素诱导的丝氨酸/苏氨酸蛋白激酶，它能磷酸化真核细胞起始因子2的α亚基，以应对病毒感染。经典的遗传学方法研究PKR在细胞信号中的作用有其局限性，这是因为重叠但非冗余的通路。PKR的小分子抑制剂在这方面将是有用的。我们在此报道，发现了一种抑制蛋白酪氨酸激酶反应的小分子抑制剂。该抑制剂是通过筛选26个不同结构的不同ATP结合位点导向的抑制剂文库而发现的。我们还描述了使用兔网织红细胞裂解物系统和荧光素酶mRNA高通量筛选PKR抑制剂的大量化合物的方法。该分析利用了网织红细胞裂解物含有丰富的翻译机制组件的事实，而PKR是其中不可或缺的一部分。本实验可以在外源性人PKR的作用下，研究不同化合物对人PKR造成的翻译障碍的修复作用。(C)2003 Elsevier Inc.保留所有权利。
The RNA-dependent protein kinase (PKR) is an interferon-induced serine/threonine protein kinase that phosphorylates the alpha subunit of the eukaryotic initiation factor 2 in response to viral infection. Classical genetic approaches for studying the role of PKR in cell signaling have their limitations due to overlapping but non-redundant pathways. Small molecule inhibitors of PKR will be useful in this regard. We report here, the discovery of a small molecule inhibitor of the kinase reaction of PKR. The inhibitor was discovered by screening a library of 26 different ATP-binding site directed inhibitors of varying structure. We also describe the development of a high-throughput assay for screening a large number of compounds for,a PKR inhibitor using a rabbit reticulocyte lysate system and luciferase mRNA. The assay takes advantage of the fact that the reticulocyte lysate is rich in components of the translational machinery, of which PKR is an integral part. This assay can be carried out with added exogenous human PKR to study the effect of various compounds in their ability to rescue the translational block imposed by human PKR. (C) 2003 Elsevier Inc. All rights reserved.