Structural basis of the allosteric inhibitor interaction on the HIV-1 reverse transcriptase RNase H domain.
Structural basis of the allosteric inhibitor interaction on the HIV-1 reverse transcriptase RNase H domain.
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DOI：
10.1111/cbdd.12010
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发表时间：
2012-11
影响因子：
3
通讯作者：
Ishima R
中科院分区：
文献类型：
--
作者：
Christen MT;Menon L;Myshakina NS;Ahn J;Parniak MA;Ishima R
HIV-1逆转录酶（RT）是抗逆转录病毒药物开发的一个重要靶点。虽然这种酶是双功能的，具有DNA聚合酶和核糖核酸酶H（RNH）活性，但没有临床批准的RNH活性抑制剂。在这里，我们描述了变构位点抑制剂BHMP 07与野生型（WT）RNH片段的结构基础和分子相互作用。WT RNH上的抑制剂滴定的溶液NMR实验显示出相对较宽的化学位移扰动，表明抑制剂相互作用的长程构象效应。比较了Mg 2+存在和不存在时RNH与RNH二聚体的NMR化学位移变化，以确定和验证相互作用位点。借助分子对接的NMR结果表明，BHMP 07优先结合位于RNH活性位点与包含螺旋B和D的区域（“底物-手柄区域”）之间的位点。相互作用位点与先前提出的使用嵌合RNH（pl 5-EC）鉴定的位点一致[Gong，el（2011）Chem. Biol. Drug Des. 77，39-47]，但与WT RNH相比，具有反映p15-EC中氨基酸序列特征的轻微差异。
HIV-1 reverse transcriptase (RT) has been an attractive target for the development of antiretroviral agents. Although this enzyme is bi-functional, having both DNA polymerase and ribonuclease H (RNH) activities, there is no clinically approved inhibitor of the RNH activity. Here, we characterize the structural basis and molecular interaction of an allosteric site inhibitor, BHMP07, with the wild type (WT) RNH fragment. Solution NMR experiments for inhibitor titration on WT RNH showed relatively wide chemical shift perturbations, suggesting a long-range conformational effect on the inhibitor interaction. Comparisons of the inhibitor-induced NMR chemical-shift changes of RNH with those of RNH dimer, in the presence and absence of Mg2+, were performed to determine and verify the interaction site. The NMR results, with assistance of molecular docking, indicate that BHMP07 preferentially binds to a site that is located between the RNH active site and the region encompassing helices B and D (the “substrate-handle region”). The interaction site is consistent with the previous proposed site, identified using a chimeric RNH (p15-EC) [Gong, el (2011) Chem. Biol. Drug Des. 77, 39-47], but with slight differences that reflect the characteristics of the amino acid sequences in p15-EC compared to the WT RNH.
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影响因子：
5.4
作者：
Lim, David;Gregorio, G. Glenn;Goff, Stephen P.
通讯作者：
Goff, Stephen P.
影响因子：
3
作者：
Gong, Qingguo;Menon, Lakshmi;Ishima, Rieko
通讯作者：
Ishima, Rieko
影响因子：
7.3
作者：
Kirschberg, Thorsten A.;Balakrishnan, Mini;Lansdon, Eric B.
通讯作者：
Lansdon, Eric B.
影响因子：
8
作者：
Kern, G;Handel, T;Marqusee, S
通讯作者：
Marqusee, S
DOI：
10.1093/protein/12.3.189
发表时间：
1999-03-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
Arico-Muendel, CC;Patera, A;Wolfson, AJ
通讯作者：
Wolfson, AJ