Background: Reverse Transcriptase (RT) of immunodeficiency virus type-1 (HIV-1) remains an essential target for new antiretroviral therapies. Non-nucleoside reverse transcriptase inhibitors (or NNRTIs) constitute a major class of RT inhibitors whose characterization is essential.

Introduction: Several biochemical, biological, and biophysical methods have been previously used to analyze the biological effects of NNRTIs. We explored here the use of surface plasmonic resonance to characterize the affinity of RT towards selected NNRTIs and compared the results with those obtained with in vitro and in cellulo assays.

Methods: The solubility and stability in buffers of the tested NNRTIs were assessed by spectrophotometry and fluorescence. Surface plasmonic resonance experiments to study direct NNRTIs binding to immobilized RT and intramolecular quenching of RT tryptophan fluorescence were used to determine the KA association constants (= 1/KD) between RT and the inhibitors. The in vitro inhibition constants of RT were determined using kinetics and the effects on three other potential targets (proteasome, HIV-1 integrase, and HIV-1 protease) were analyzed.

Results: The results obtained with two typical molecules belonging to our previous N-hydroxyureido acylnucleoside derivatives series using the above biophysical assays matched those obtained in in vitro and previous in cellulo assays.

Conclusion: Surface plasmonic resonance provides reliable thermodynamic information on the interaction of RT with NNRTIs and appears as a useful method for understanding their inhibitory mechanism.

背景：1型免疫缺陷病毒（HIV-1）的逆转录酶（RT）仍然是新抗逆转录病毒疗法的重要目标。非核苷逆转录酶抑制剂（或NNRTIs）构成了一类主要的RT抑制剂，其表征至关重要。

简介：以前已经使用了几种生化，生物学和生物物理方法来分析NNRTI的生物学效应。我们在这里探索了使用表面等离子体共振来表征RT对选定的NNRTIs的亲和力，并将结果与​​体外和纤维素分析中获得的结果进行了比较。

方法：通过分光光度法和荧光法评估所测试的NNRTI在缓冲液中的溶解度和稳定性。表面等离子共振实验用于研究直接NNRTIs与固定化RT的直接结合以及RT色氨酸荧光的分子内猝灭，用于确定RT与抑制剂之间的KA关联常数（= 1 / KD）。使用动力学确定RT的体外抑制常数，并分析对其他三种潜在靶标（蛋白酶体，HIV-1整合酶和HIV-1蛋白酶）的影响。

结果：使用上述生物物理测定法，使用属于我们先前的N-羟基脲基酰基核苷衍生物系列的两个典型分子所获得的结果与体外和先前在纤维素测定法中获得的结果相匹配。

结论：表面等离子共振为RT与NNRTIs的相互作用提供了可靠的热力学信息，似乎是了解其抑制机理的有用方法。