title: Screening for HIV protease inhibitors by protection against activity-mediated cytotoxicity in Escherichia coli.
abstract: Expressed retroviral proteases are often cytotoxic to the hosts. The cytotoxicity of a tethered dimer HIV protease described previously is particularly severe that transformed Escherichia coli cells could not survive the bactericidal activity of the low-level protease produced under uninduced conditions. The presence of HIV protease inhibitors protected the transformed cells from cytotoxic effects and allowed the growth of these cells on plates and in broth. A high throughput screening method was developed to seek compounds that served as "growth factors" for the HIV protease restricted cells. Several compounds identified by this screening supported the growth of these cells, preserved their viability, and inhibited HIV protease. This assay could be used as a general method for screening for inhibitors of recombinant enzymes that produce a cytotoxic phenotype in host cells.
authors.affiliation: Genomics Research Center, Academia Sinica, 128 Academia Road, Section 2, Nankang, Taipei 115, Taiwan, ROC. ysecheng@gate.sinica.edu.tw
authors.forename: Yih-Shyun E
authors.initials: YS
authors.lastname: Cheng
authors.forename: Kien-Hock
authors.initials: KH
authors.lastname: Lo
authors.forename: Hsien-Hua
authors.initials: HH
authors.lastname: Hsu
authors.forename: Yi-Ming
authors.initials: YM
authors.lastname: Shao
authors.forename: Wen-Bin
authors.initials: WB
authors.lastname: Yang
authors.forename: Chun-Hung
authors.initials: CH
authors.lastname: Lin
authors.forename: Chi-Huey
authors.initials: CH
authors.lastname: Wong
mesh_terms: D015169:Colony Count, Microbial; D003720:Densitometry; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D015964:Gene Expression Regulation, Bacterial; D016333:HIV Protease / Q000187:drug effects* / Q000235:genetics / Q000633:toxicity*; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000187:drug effects / Q000235:genetics / Q000633:toxicity*; D014169:Transformation, Bacterial
chemical_list: D017320:HIV Protease Inhibitors; D011994:Recombinant Proteins; D016333:HIV Protease