Development And Application Of SPR-based Assays For Screening HIV-1Vpu Inhibitors From Small Molecular Compounds

Acquired Immunodeficiency Syndrom (AIDS), is caused by humanimmunodeficiency virus (HIV) infection causes chronic disease. Since there has beenno complete cure for AIDS drugs available, therefore, it is important to find promisinganti-HIV drugs.Viral protein U (Vpu), a trans-membrane accessory protein encoded by HIV-1, isknown to enhance the release of newly formed virions and is becoming a promisingtarget for development of anti-HIV drugs. However, the drug screening methods oftargeted Vpu protein are limited, the development of target drug is only in the earlystage. Therefore, it is significant that establish a new drug screening method of targetfor Vpu protein.This study is arm to establish a drug screening method of Vpu protein by SPRtechnology. The methods including the Pichia pastoris expression system was used toexpress and purify the Vpu protein. The recombinant Vpu protein were then attachedto the surface of BIAcoreT MCM5biosensor chip for screening of high-affinity anti-Vpu drugs from small molecular compounds. Furthermore,293FT cells or THN-positive cells were transfected with pEGFP-vphu plasmid/Halotag-THN plasmid andexamined for Vpu and THN expression after compounds treatment and the cytotoxic ityof positive compounds using cell experiments. In addition, MAGI tests were applied toidentify the in vitro anti-HIV effects of positive compounds.The results were shown that three out of53herbal compounds were identified inBIAcore assay as high-affinity anti-Vpu compounds. In addition, compound#1showeda significant anti-HIV effect that was at least partly due to the restoration of Vpu-dependented HIV release function. Compound#2and#3also exhibited strong anti-HIV effects, however the mechanisms were not related to Vpu.In all, the SPR-based model has been successfully used in screening anti-Vpuinhibitors in our study and will contribute to the future discovery of effective anti-HIVdrugs.