pH-induced destabilization of lipid bilayers by a lipopeptide derived from influenza hemagglutinin.
A synthetic twenty-one amino acid peptide (AcE4K) based on the amino acid sequence of the influenza HA2 fusion peptide was coupled to a distearoylglycerol lipid anchor by amidation of an N-terminal lysine side chain. The secondary structure of Lipo-AcE4K incorporated into POPC (1-palmitoyl-2-oleoyl-sn-phosphatidylcholine) liposomes was not measurably affected by pH, but increased membrane penetration was indicated by tryptophan fluorescence. At outer monolayer concentrations up to 10 mol%, Lipo-AcE4K formed stable liposomes with POPC and EPC/Chol (egg phosphatidylcholine/cholesterol) (55:45) at pH 7.5. Acid-induced destabilization and fusion of these vesicles were demonstrated by fluorescent lipid mixing and contents leakage assays, and by freeze-fracture electron microscopy. Membrane destabilization increased with increasing lipopeptide concentrations, decreasing pH, inclusion of cholesterol, and incorporation of lipopeptide into the inner monolayer as well as the outer monolayer of the liposomes. Fusion of liposomes bearing Lipo-AcE4K with erythrocyte ghosts was demonstrated by lipid mixing and fluorescence microscopy.
Bailey
A L
AL
Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, Canada. 71612.771@compuserve.com
Monck
M A
MA
Cullis
P R
PR
D000595:Amino Acid Sequence; D002784:Cholesterol / Q000737:chemistry; D004910:Erythrocyte Membrane / Q000737:chemistry; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008051:Lipid Bilayers / Q000737:chemistry*; D008074:Lipoproteins / Q000737:chemistry; D008081:Liposomes; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry*; D017433:Protein Structure, Secondary; D012995:Solubility
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D008051:Lipid Bilayers; D008074:Lipoproteins; D008081:Liposomes; D010455:Peptides; D010713:Phosphatidylcholines; D002784:Cholesterol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine