A sensitive HIV-1 envelope induced fusion assay identifies fusion enhancement of thrombin

De Chun Cheng, Guo Cai Zhong, Ju Xiang Su, Yan Hong Liu, Yan Li, Jia Ye Wang, Toshio Hattori, Hong Ling, Feng Min Zhang

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

To evaluate the interaction between HIV-1 envelope glycoprotein (Env) and target cell receptors, various cell-cell-fusion assays have been developed. In the present study, we established a novel fusion system. In this system, the expression of the sensitive reporter gene, firefly luciferase (FL) gene, in the target cells was used to evaluate cell fusion event. Simultaneously, constitutively expressed Renilla luciferase (RL) gene was used to monitor effector cell number and viability. FL gave a wider dynamic range than other known reporters and the introduction of RL made the assay accurate and reproducible. This system is especially beneficial for investigation of potential entry-influencing agents, for its power of ruling out the false inhibition or enhancement caused by the artificial cell-number variation. As a case study, we applied this fusion system to observe the effect of a serine protease, thrombin, on HIV Env-mediated cell-cell fusion and have found the fusion enhancement activity of thrombin over two R5-tropic HIV strains.

Original languageEnglish
Pages (from-to)1780-1784
Number of pages5
JournalBiochemical and biophysical research communications
Volume391
Issue number4
DOIs
Publication statusPublished - 2010 Jan 22

Keywords

  • Cell fusion
  • Dual-reporter
  • Envelope
  • HIV-1
  • Thrombin

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Cheng, D. C., Zhong, G. C., Su, J. X., Liu, Y. H., Li, Y., Wang, J. Y., Hattori, T., Ling, H., & Zhang, F. M. (2010). A sensitive HIV-1 envelope induced fusion assay identifies fusion enhancement of thrombin. Biochemical and biophysical research communications, 391(4), 1780-1784. https://doi.org/10.1016/j.bbrc.2009.12.155