A mammalian two-hybrid system-based assay for small-molecular HIV fusion inhibitors targeting gp41

作者：Shui Xiaoxi; Lu Xinhua; Gao Yan; Liu Chen; Ren Fengzhi; Jiang Qin; Zhang Hua; Zhao Baohua^*; Zheng Zhihui

来源：Antiviral Research, 2011, 90(1): 54-63.

DOI：10.1016/j.antiviral.2011.02.006

• HIV fusion inhibitor
• gp41
• Mammalian two-hybrid system
• Small molecular compound

gp41 is a major component of the envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1) responsible for fusion of the viral envelope with the target cellular membrane. The formation of the trimer-of-hairpins core structure of gp41, via the interaction between its N-terminal heptad repeat (NHR) and its C-terminal heptad repeat (CHR) plays a key role in the membrane fusion process. Hence, inhibitors of trimer-of-hairpins formation have become a promising new class of HIV therapeutics. In the present study, based on the mammalian two-hybrid system, we developed a cell-based assay for detecting small-molecular HIV-1 fusion inhibitors targeting gp41. The optimized assay can be adapted to high-throughput screening in 96- and 384-well microplates with high signal-to-background ratios and acceptable Z' factors. The known small-molecular gp41 inhibitors, ADS-it, XTT formazan and tannin acid, tested positive in this assay, with half-maximal inhibitory concentration (IC(50)) values of 4.9 mu M, 5.6 mu M and 0.8 mu M, respectively. These data suggested that this novel assay is robust, sensitive and specific for identifying small-molecular HIV-1 gp41 inhibitors.

• 出版日期2011-4
• 单位河北师范大学