The polar region consecutive to the HIV fusion peptide participates in membrane fusion.
The fusion peptide of HIV-1 gp41 is formed by the 16 N-terminal residues of the protein. This 16-amino acid peptide, in common with several other viral fusion peptides, caused a reduction in the bilayer to hexagonal phase transition temperature of dipalmitoleoylphosphatidylethanolamine (T(H)), suggesting its ability to promote negative curvature in membranes. Surprisingly, an elongated peptide corresponding to the 33 N-terminal amino acids raised T(H), although it was more potent than the 16-amino acid fusion peptide in inducing lipid mixing with large unilamellar liposomes of 1:1:1 dioleoylphosphatidylethanolamine/dioleoylphosphatidylcholine/choleste rol. The 17-amino acid C-terminal fragment of the peptide can induce membrane fusion by itself, if it is anchored to a membrane by palmitoylation of the amino terminus, indicating that the additional 17 hydrophilic amino acids contribute to the fusogenic potency of the peptide. This is not solely a consequence of the palmitoylation, as a random peptide with the same amino acid composition with a palmitoyl anchor was less potent in promoting membrane fusion and palmitic acid itself had no fusogenic activity. The 16-amino acid N-terminal fusion peptide and the longer 33-amino acid peptide were labeled with NBD. Fluorescence binding studies indicate that both peptides bind to the membrane with similar affinities, indicating that the increased fusogenic activity of the longer peptide was not a consequence of a greater extent of membrane partitioning. We also determined the secondary structure of the peptides using FTIR spectroscopy. We find that the amino-terminal fusion peptide is inserted into the membrane as a beta-sheet and the 17 C-terminal amino acids lie on the surface of the membrane, adopting an alpha-helical conformation. It was further demonstrated with the use of rhodamine-labeled peptides that the 33-amino acid peptide self-associated in the membrane while the 16-amino acid N-terminal peptide did not. Thus, the 16-amino acid N-terminal fusion peptide of HIV inserts into the membrane and, like other viral fusion peptides, lowers T(H). In addition, the 17 consecutive amino acids enhance the fusogenic activity of the fusion peptide presumably by promoting its self-association.
Peisajovich
S G
SG
Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
Epand
R F
RF
Pritsker
M
M
Shai
Y
Y
Epand
R M
RM
D000595:Amino Acid Sequence; D015700:HIV Envelope Protein gp41 / Q000737:chemistry* / Q000502:physiology; D015497:HIV-1 / Q000502:physiology; D006801:Humans; D008051:Lipid Bilayers / Q000737:chemistry; D008081:Liposomes; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D010169:Palmitic Acids / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry* / Q000502:physiology; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D014760:Viral Fusion Proteins / Q000737:chemistry* / Q000502:physiology; D019065:Virus Assembly
D015700:HIV Envelope Protein gp41; D008051:Lipid Bilayers; D008081:Liposomes; D010169:Palmitic Acids; D010446:Peptide Fragments; D014760:Viral Fusion Proteins