A novel enzyme-linked immunosorbent assay for screening HIV-1 fusion inhibitors targeting HIV-1 Gp41 core structure

• Published in: Journal of biomolecular screening Vol. 16; no. 2; pp. 221 - 229
• Main Authors: Pang, Wei, Wang, Rui-Rui, Gao, Yue-Dong, Yang, Liu-Meng, Sun, Yi, Huang, Jing-Fei, Tien, Po, Zheng, Yong-Tang
• Format: Journal Article
• Language: English
• Published: United States 01.02.2011
• Subjects: Amino Acid Sequence; Anti-HIV Agents - metabolism; Anti-HIV Agents - pharmacology; Cell Line; Drug Evaluation, Preclinical; Enzyme-Linked Immunosorbent Assay - methods; Giant Cells - drug effects; HIV Envelope Protein gp41 - chemistry; HIV Envelope Protein gp41 - metabolism; HIV Fusion Inhibitors - metabolism; HIV Fusion Inhibitors - pharmacology; HIV-1 - drug effects; Humans; Molecular Sequence Data; Peptides - metabolism; Protein Structure, Secondary; Recombinant Fusion Proteins - metabolism; Reproducibility of Results; Sensitivity and Specificity
• ISSN: 2472-5552; 1552-454X
• DOI: 10.1177/1087057110393333

The gp41 subunit of the human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein mediates the fusion of viral and host cell membranes. As the HIV-1 enters the host cells, the 2 helical regions, HR1 and HR2, in the ectodomain of gp41 can form a 6-helix bundle, which brings the viral and target cell membranes to close proximity and serves as an attractive target for developing HIV-1 fusion inhibitors. Now, there are several cell- and molecule-based assays to identify potential HIV-1 fusion inhibitors targeting gp41. However, these assays cannot be used universally because they are time-consuming, inconvenient, and expensive. In the present study, the authors expressed and purified GST-HR121 and C43-30a proteins that were derived from the HIV-1 gp41 ectodomain region. GST-HR121 has a function similar to the HR1 peptide of gp41, whereas C43-30a is an HR2-derived peptide that added 50 amino acid residues (aa) in the N-terminal of C43. Further research found they could interact with each other, and a potential HIV-1 fusion inhibitor could inhibit this interaction. On the basis of this fact, a novel, rapid, and economic enzyme-linked immunosorbent assay was established, which can be developed for high-throughput screening of HIV-1 fusion inhibitors.