Monitoring of HIV-1 envelope-mediated membrane fusion using modified split green fluorescent proteins.
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.
Wang
Jianqi
J
China-Japan Joint Laboratory of Structural Virology and Immunology, Institute of Biophysics, Chinese Academy of Sciences, Chaoyang District, Beijing, China.
Kondo
Naoyuki
N
Long
Yufei
Y
Iwamoto
Aikichi
A
Matsuda
Zene
Z
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D049452:Green Fluorescent Proteins; D015700:HIV Envelope Protein gp41 / Q000008:administration & dosage; D023581:HIV Fusion Inhibitors / Q000008:administration & dosage; D015658:HIV Infections / Q000821:virology*; D015497:HIV-1 / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008561:Membrane Fusion / Q000187:drug effects*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000008:administration & dosage; D014773:Virology / Q000379:methods; D054299:env Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*
D015700:HIV Envelope Protein gp41; D023581:HIV Fusion Inhibitors; D010446:Peptide Fragments; D054299:env Gene Products, Human Immunodeficiency Virus; C407149:peptide C34; D049452:Green Fluorescent Proteins