Synthesis, enhanced fusogenicity, and solid state NMR measurements of cross-linked HIV-1 fusion peptides.
In the HIV-1 gp41 and other viral fusion proteins, the minimal oligomerization state is believed to be trimeric with three N-terminal fusion peptides inserting into the membrane in close proximity. Previous studies have demonstrated that the fusion peptide by itself serves as a useful model fusion system, at least to the hemifusion stage in which the viral and target cell lipids are mixed. In the present study, HIV-1 fusion peptides were chemically synthesized and cross-linked at their C-termini to form dimers or trimers. C-terminal trimerization is their likely topology in the fusogenic form of the intact gp41 protein. The fusogenicity of the peptides was then measured in an intervesicle lipid mixing assay, and the assay results were compared to those of the monomer. For monomer, dimer, and trimer at peptide strand/lipid mol ratios between 0.0050 and 0.010, the final extent of lipid mixing for the dimer and trimer was 2-3 times greater than for the monomer. These data suggest that the higher local concentration of peptide strands in the cross-linked peptides enhances fusogenicity and that oligomerization of the fusion peptide in gp41 may enhance the rate of viral/target cell membrane fusion. For gp41, this effect is in addition to the role of the trimeric coiled-coil structure in bringing about apposition of viral and target cell membranes. NMR measurements on the membrane-associated dimeric fusion peptide were consistent with an extended structure at Phe-8, which is the same as has been observed for the membrane-bound monomer in the same lipid composition.
Yang
Rong
R
Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, USA.
Yang
Jun
J
Weliky
David P
DP
D000595:Amino Acid Sequence; D003432:Cross-Linking Reagents; D019281:Dimerization; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000502:physiology; D015497:HIV-1; D006801:Humans; D066298:In Vitro Techniques; D008055:Lipids; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D020836:Protein Structure, Quaternary; D014760:Viral Fusion Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000502:physiology
D003432:Cross-Linking Reagents; D015700:HIV Envelope Protein gp41; D008055:Lipids; D014760:Viral Fusion Proteins