Development of a fluorescence-based HIV-1 integrase DNA binding assay for identification of novel HIV-1 integrase inhibitors

Ying Shan Han, Wei Lie Xiao, Peter K. Quashie, Thibault Mesplède, Hongtao Xu, Eric Deprez, Olivier Delelis, Jian Xin Pu, Han Dong Sun, Mark A. Wainberg

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

Human immunodeficiency virus integrase (HIV-1 IN) inhibitors that are currently approved or are in advanced clinical trials specifically target the strand transfer step of integration. However, considerable cross-resistance exists among some members of this class of IN inhibitors. Intriguingly, though, HIV-1 IN possesses multiple sites, distinct from those involved in the strand transfer step, that could be targeted to develop new HIV-1 IN inhibitors. We have developed a fluorescent HIV-1 IN DNA binding assay that can identify small molecules termed IN binding inhibitors (INBIs) that inhibit IN binding to viral DNA. This assay has been optimized with respect to concentrations of each protein, long terminal repeat (LTR) DNA substrate, salt, and time, and has been used successfully to measure the HIV-1 IN DNA binding activity of a well-characterized INBI termed FZ41. In addition, we have used the assay to screen a small library of natural products, resulting in the identification of nigranoic acid as a new INBI. The proposed fluorescence assay is easy and inexpensive, and provides a high-throughput detection method for determination of HIV-1 IN DNA binding activity, monitoring of enzyme kinetics, and high-throughput screening for the identification of new INBIs.

Original languageEnglish
Pages (from-to)441-448
Number of pages8
JournalAntiviral Research
Volume98
Issue number3
DOIs
Publication statusPublished - Jun 2013
Externally publishedYes

Keywords

  • Fluorescent assay
  • HIV-1 integrase inhibitors
  • Integrase DNA binding

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