NMR characterization of HIV-1 reverse transcriptase binding to various non-nucleoside reverse transcriptase inhibitors with different activities.
NMR characterization of HIV-1 reverse transcriptase binding to various non-nucleoside reverse transcriptase inhibitors with different activities.
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DOI：
10.1038/srep15806
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发表时间：
2015-10-29
影响因子：
4.6
通讯作者：
Hannongbua S
中科院分区：
文献类型：
--
作者：
Thammaporn R;Yagi-Utsumi M;Yamaguchi T;Boonsri P;Saparpakorn P;Choowongkomon K;Techasakul S;Kato K;Hannongbua S
人类免疫缺陷病毒1型逆转录酶（HIV-1 RT）是获得性免疫缺陷综合征抗病毒治疗的重要靶点。然而，现有药物的效率受损最典型的是这种酶的耐药性突变。在这项研究中，我们应用核磁共振（NMR）光谱技术来表征HIV-1 RT与各种具有不同活性的非核苷类逆转录酶抑制剂（NNRTIs）的结合，即奈韦拉平、德拉韦林、依非韦伦、达匹韦林、伊曲维林和利匹韦林。在存在和不存在这些NNRTIs的情况下，收集HIV-1 RT的h -13C异核单量子相干（HSQC）光谱数据，其中p66亚基的蛋氨酸甲基被选择性地标记为13C。我们发现这些药物结合的HIV-1 RT的M230共振的甲基13C化学位移与它们的抗HIV-1 RT活性高度相关。这种蛋氨酸残基位于nnrti结合口袋附近，但不直接参与药物相互作用，而是作为构象探针，表明HIV-1 RT的开放构象中更多地填充了具有更高抑制活性的nnrti。因此，核磁共振方法为开发抗hiv药物筛选新型nnrti提供了一个有用的工具。
Human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) is an important target for antiviral therapy against acquired immunodeficiency syndrome. However, the efficiency of available drugs is impaired most typically by drug-resistance mutations in this enzyme. In this study, we applied a nuclear magnetic resonance (NMR) spectroscopic technique to the characterization of the binding of HIV-1 RT to various non-nucleoside reverse transcriptase inhibitors (NNRTIs) with different activities, i.e., nevirapine, delavirdine, efavirenz, dapivirine, etravirine, and rilpivirine. 1H-13C heteronuclear single-quantum coherence (HSQC) spectral data of HIV-1 RT, in which the methionine methyl groups of the p66 subunit were selectively labeled with 13C, were collected in the presence and absence of these NNRTIs. We found that the methyl 13C chemical shifts of the M230 resonance of HIV-1 RT bound to these drugs exhibited a high correlation with their anti-HIV-1 RT activities. This methionine residue is located in proximity to the NNRTI-binding pocket but not directly involved in drug interactions and serves as a conformational probe, indicating that the open conformation of HIV-1 RT was more populated with NNRTIs with higher inhibitory activities. Thus, the NMR approach offers a useful tool to screen for novel NNRTIs in developing anti-HIV drugs.