Structure of HIV-1 Reverse Transcriptase with the Inhibitor β-Thujaplicinol Bound at the RNase H Active Site
Structure of HIV-1 Reverse Transcriptase with the Inhibitor β-Thujaplicinol Bound at the RNase H Active Site
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DOI：
10.1016/j.str.2009.09.016
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发表时间：
2009-12-09
期刊：
影响因子：
5.7
通讯作者：
Arnold, Eddy
中科院分区：
文献类型：
--
作者：
Himmel, Daniel M.;Maegley, Karen A.;Arnold, Eddy
需要新的抑制剂来对抗抗药性艾滋病毒变种的迅速出现。HIV-1逆转录酶(RT)同时具有DNA聚合酶和RNase H(RNH)酶活性，但已批准的抑制RT的药物针对该聚合酶。针对新靶点的抑制剂，如RNH，应该对所有当前的耐药变异体有效。在这里，我们给出了一种RNH抑制剂--β-胡萝卜素的2.80埃和2.04埃分辨率的晶体结构，它结合在HIV-1RT的RNH活性部位和一个分离的RNH结构域上。β-胡萝卜素在RNH活性部位螯合两个二价金属离子。我们提供的生化证据表明，β-胡黄素是一种慢结合的RNH抑制剂，具有非竞争性动力学，并表明它形成了一种与RT和RNA：DNA底物有利相互作用的三联吡啶离子。
Novel inhibitors are needed to counteract the rapid emergence of drug-resistant HIV variants. HIV-1 reverse transcriptase (RT) has both DNA polymerase and RNase H (RNH) enzymatic activities, but approved drugs that inhibit RT target the polymerase. Inhibitors that act against new targets, such as RNH, should be effective against all of the current drug-resistant variants. Here, we present 2.80 angstrom and 2.04 angstrom resolution crystal structures of an RNH inhibitor, beta-thujaplicinol, bound at the RNH active site of both HIV-1 RT and an isolated RNH domain. beta-thujaplicinol chelates two divalent metal ions at the RNH active site. We provide biochemical evidence that beta-thujaplicinol is a slow-binding RNH inhibitor with noncompetitive kinetics and suggest that it forms a tropylium ion that interacts favorably with RT and the RNA:DNA substrate.