Human immunodeficiency virus (HIV)-1 reverse transcriptase (HIV-1 RT) is responsible for the transcription of viral RNA genomes into DNA genomes and has become an important target for the treatment of acquired immune deficiency syndrome (AIDS). This study used biophysical techniques to characterize the HIV-1 RT structure, monomer forms, and the non-nucleoside reverse transcriptase inhibitors (NNRTIs) bound forms. Inactive p66_W401A and p51_W401A were selected as models to study the HIV-1 RT monomer structures. Nuclear magnetic resonance (NMR) spectroscopy revealed that the unliganded forms of p66_W401A protein and p51_W401A protein had similar conformation to each other in solution. The complexes of p66_W401A or p51_W401A with inhibitors showed similar conformations to p66 in the RT heterodimer bound to the NNRTIs. Furthermore, the results of paramagnetic relaxation enhancement (PRE)-assisted NMR revealed that the unliganded forms of the p66_W401A and p51_W401A conformations were different from the unliganded heterodimer, characterized by a greater distance between the fingers and thumb subdomains. Small-angle X-ray scattering (SAXS) experiments confirmed that p66_W401A and p51_W401A can bind with inhibitors, similar to the p66/p51 heterodimer. The findings of this study increase the structural knowledge base of HIV-1 RT monomers, which may be helpful in the future design of potent viral inhibitors.

人类免疫缺陷病毒（HIV）-1逆转录酶（HIV-1 RT）负责将病毒RNA基因组转录为DNA基因组，已成为治疗获得性免疫缺陷综合征（AIDS）的重要靶点。本研究使用生物物理技术来表征 HIV-1 RT 结构、单体形式和非核苷逆转录酶抑制剂 (NNRTI) 结合形式。选择失活的p66 _W401A和p51 _{W401A作为模型来研究HIV-1 RT单体结构。}核磁共振（NMR）光谱显示p66 _W401A蛋白和p51 _W401A蛋白的未配体形式在溶液中具有相似的构象。_{p66 W401A}或 p51 _W401A与抑制剂的复合物显示出与与 NNRTI 结合的 RT 异二聚体中的 p66 相似的构象。此外，顺磁弛豫增强（PRE）辅助NMR的结果表明，p66 _W401A和p51 _W401A构象的未配体形式与未配体异二聚体不同，其特征在于手指和拇指子结构域之间的距离更大。小角X射线散射（SAXS）实验证实p66 _W401A和p51 _W401A可以与抑制剂结合，类似于p66/p51异二聚体。这项研究的结果增加了 HIV-1 RT 单体的结构知识库，这可能有助于未来有效病毒抑制剂的设计。