Vpu蛋白是HIV-1病毒特有的与病毒释放相关的辅助蛋白,正成为抗HIV药物研究的热点。本项目以Vpu蛋白为靶点，利用"非标记"的SPR技术构建Vpu互作分子筛选模型，针对Vpu蛋白不同结构域进行抑制剂的发现并研究其作用机制。项目利用原核细胞表达Vpu蛋白及片段，利用氨基偶联、His-tag捕获等方式将蛋白固定在芯片表面，通过小分子化合物与Vpu蛋白的相互作用进行整体水平的亲和力评价；利用化合物与Vpu跨膜区和胞质区相互作用的差异进行结合位点的分析；选取与Tetherin蛋白竞争结合的候选物、Vpu跨膜区候选物及Vpu胞质区候选物进行细胞水平抗HIV-1研究。结合小分子化合物与Vpu蛋白相互作用情况及抗HIV活性结果进行构效关系分析和作用机制研究，探索Vpu蛋白不同功能区与抗HIV-1作用的相关性，并最终从候选物中发现高效低毒的抗HIV先导物，为开发具有自主知识产权的抗HIV新药奠定基础。

Vpu protein is one of special HIV-1 virus accessory proteins and related to HIV-1 release. It's becoming a hotspot for anti-HIV research. In this project, an interaction screening model targeted the HIV-1 Vpu protein will be developed by "label-free" SPR technology, and the inhibitors targeted the various structural and functional domain will be found. Vpu protein and its functional segments which are expressed and purified from E.coli cell will be immobilized on the sensor chip surface by amino coupling or His-tag capture. The affinity of the interaction between the small molecule compounds and Vpu protein will be evaluated. The binding sites for small molecule compounds will be confirmed by analysis the difference between the interaction with Vpu protein transmembrane domain and cytoplasmic segment. Three candidates which are competing with Tetherin protein, binding with Vpu transmembrane domain and binding with cytoplasmic segment will be selected to research anti-HIV activity on the cell level. According to the interaction between compounds and Vpu protein and the results of anti-HIV activity, the structure-activity relationship and mechanism will be announced. The relationship of the different domain of Vpu protein and anti-HIV activity will be explored. Some effective and low toxicity anti-HIV lead compounds of anti-HIV drugs will be found and developed.