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毛细管电泳法研究 HIV 逆转录酶与其非核苷抑制剂奈韦拉平的相互作用。

Study on the interaction between HIV reverse transcriptase and its non-nucleoside inhibitor nevirapine by capillary electrophoresis.

机构信息

Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Jun 15;878(20):1714-7. doi: 10.1016/j.jchromb.2010.04.013. Epub 2010 Apr 24.

Abstract

The HIV reverse transcriptase (RT) is an important antiviral target for the chemotherapy of AIDS because of its key role in virus replication. Nevirapine is a first generation of non-nucleoside reverse transcriptase inhibitors (NNRTIs), which is usually used for the therapy of AIDS. In this study, a high-performance analytical method based on capillary electrophoresis (CE) to investigate interactions between HIV RT and nevirapine was developed. Samples containing HIV RT and nevirapine at various ratios were incubated at 37 degrees C for 45 min and then separated by CE with Tris-acetate buffer at pH 7.3 containing 0.15% SDS. Both qualitative and quantitative characterizations of the binding were determined by CE for the first time. The binding constants of the interactions between HIV RT and nevirapine were calculated as (3.25+/-0.16)x10(4) and (1.25+/-0.07)x10(2) M(-1) by Scatchard analysis. HIV RT and nevirapine have two binding sites. The presented methodology should be generally applicable to study the interactions between HIV RT and nevirapine quantitatively and qualitatively.

摘要

人类免疫缺陷病毒(HIV)逆转录酶(RT)在病毒复制中起着关键作用,是抗艾滋病化疗的重要抗病毒靶标。奈韦拉平是第一代非核苷类逆转录酶抑制剂(NNRTIs),通常用于艾滋病的治疗。在这项研究中,建立了一种基于毛细管电泳(CE)的高效分析方法,用于研究 HIV RT 与奈韦拉平之间的相互作用。将含有不同比例 HIV RT 和奈韦拉平的样品在 37°C 下孵育 45 分钟,然后用 pH 值为 7.3 的 Tris-醋酸盐缓冲液分离,其中含有 0.15% SDS。首次通过 CE 对结合进行了定性和定量表征。通过 Scatchard 分析,计算出 HIV RT 与奈韦拉平相互作用的结合常数分别为(3.25+/-0.16)x10(4)和(1.25+/-0.07)x10(2)M(-1)。HIV RT 和奈韦拉平有两个结合位点。所提出的方法应该普遍适用于定量和定性地研究 HIV RT 与奈韦拉平之间的相互作用。

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