Monitoring of HIV-1 Envelope-Mediated Membrane Fusion Using Modified Split Green Fluorescent Proteins

J Virol Methods. 2009 Nov;161(2):216-22. doi: 10.1016/j.jviromet.2009.06.017. Epub 2009 Jun 25.

Abstract

A simple, cell-based, membrane fusion assay system that uses split green fluorescent proteins (spGFPs) as an indicator was developed. The attachment of the pleckstrin homology (PH) domain to the N-termini of each spGFP not only localized the reporter signal to the plasma membrane but also helped the stable expression of the smaller spGFP of seventeen amino acid residues. It was shown that this system allowed real-time monitoring of membrane fusion by HIV-1 envelope protein (Env) without the addition of external substrates. This method can be adapted to the analyses of other viral membrane fusion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cell Line
  • Green Fluorescent Proteins
  • HIV Envelope Protein gp41 / administration & dosage
  • HIV Fusion Inhibitors / administration & dosage
  • HIV Infections / virology*
  • HIV-1 / drug effects
  • HIV-1 / metabolism*
  • Humans
  • Membrane Fusion / drug effects*
  • Molecular Sequence Data
  • Peptide Fragments / administration & dosage
  • Virology / methods
  • env Gene Products, Human Immunodeficiency Virus / metabolism*

Substances

  • HIV Envelope Protein gp41
  • HIV Fusion Inhibitors
  • Peptide Fragments
  • env Gene Products, Human Immunodeficiency Virus
  • peptide C34
  • Green Fluorescent Proteins