Interaction of HIV-1 Reverse Transcriptase Ribonuclease H with an Acylhydrazone Inhibitor
Interaction of HIV-1 Reverse Transcriptase Ribonuclease H with an Acylhydrazone Inhibitor
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DOI：
10.1111/j.1747-0285.2010.01052.x
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发表时间：
2011-01-01
影响因子：
3
通讯作者：
Ishima, Rieko
中科院分区：
文献类型：
--
作者：
Gong, Qingguo;Menon, Lakshmi;Ishima, Rieko
HIV-1逆转录酶是一种双功能酶，具有DNA聚合酶(依赖于RNA和DNA)和核糖核酸酶H的活性。HIV-1逆转录酶已经成为抗逆转录病毒治疗开发的一个非常重要的靶点，目前临床上使用的抗逆转录病毒药物中有近一半针对逆转录酶DNA聚合酶。然而，目前还没有逆转录酶核糖核酸酶H的抑制剂上市或处于临床前开发阶段。已有几种逆转录酶核糖核酸酶H的类药物小分子抑制剂被描述，但关于逆转录酶核糖核酸酶H与具有抗病毒活性的抑制剂之间的相互作用的结构信息很少。在这份报告中，我们描述了一种新的核糖核酸酶H抑制剂BHMP07与具有催化活性的HIV-1逆转录酶核糖核酸酶H结构域片段相互作用的核磁共振研究。我们进行了溶液核磁共振实验来确定BHMP07与核糖核酸酶H结构域片段的相互作用界面。不同浓度BHMP07主链酰胺信号的化学位移变化清楚地表明，BHMP07主要识别核糖核酸酶H片段中的底物手柄区域。利用核糖核酸酶H抑制试验和逆转录酶突变体，比较了BHMP07与另一种抑制剂二羟基苯甲酰肼的结合专一性。我们的结果提供了核糖核酸酶H抑制剂相互作用的结构特征，并可能有助于进一步改进抑制剂。
HIV-1 reverse transcriptase is a bifunctional enzyme, having both DNA polymerase (RNA- and DNA-dependent) and ribonuclease H activities. HIV-1 reverse transcriptase has been an exceptionally important target for antiretroviral therapeutic development, and nearly half of the current clinically used antiretrovirals target reverse transcriptase DNA polymerase. However, no inhibitors of reverse transcriptase ribonuclease H are on the market or in preclinical development. Several drug-like small molecule inhibitors of reverse transcriptase ribonuclease H have been described, but little structural information is available about the interactions between reverse transcriptase ribonuclease H and inhibitors that exhibit antiviral activity. In this report, we describe NMR studies of the interaction of a new ribonuclease H inhibitor, BHMP07, with a catalytically active HIV-1 reverse transcriptase ribonuclease H domain fragment. We carried out solution NMR experiments to identify the interaction interface of BHMP07 with the ribonuclease H domain fragment. Chemical shift changes of backbone amide signals at different BHMP07 concentrations clearly demonstrate that BHMP07 mainly recognizes the substrate handle region in the ribonuclease H fragment. Using ribonuclease H inhibition assays and reverse transcriptase mutants, the binding specificity of BHMP07 was compared with another inhibitor, dihydroxy benzoyl naphthyl hydrazone. Our results provide a structural characterization of the ribonuclease H inhibitor interaction and are likely to be useful for further improvements of the inhibitors.