Study on the interaction between HIV reverse transcriptase and its non-nucleoside inhibitor nevirapine by capillary electrophoresis.
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.
Chen
Wenjing
W
Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
Li
Wujie
W
Ling
Xiaomei
X
Wang
Xiaowei
X
Liu
Junyi
J
D019075:Electrophoresis, Capillary / Q000379:methods*; D054303:HIV Reverse Transcriptase / Q000737:chemistry*; D007700:Kinetics; D019829:Nevirapine / Q000737:chemistry*; D011485:Protein Binding; D018894:Reverse Transcriptase Inhibitors / Q000737:chemistry*
D018894:Reverse Transcriptase Inhibitors; D019829:Nevirapine; D054303:HIV Reverse Transcriptase