Interaction of a synthetic peptide corresponding to the N-terminus of canine distemper virus fusion protein with phospholipid vesicles: a biophysical study.
The F protein of canine distemper virus (CDV) is a classic type I glycoprotein formed by two polypeptides, F1 and F2. The N-terminal regions of the F1 polypeptides of CDV, measles virus and other paramyxoviruses present moderate to high homology, supporting the existence of a high conservation within these structures, which emphasises its role in viral-host cell membrane fusion. This N-terminal polypeptide is usually termed the fusion peptide. The fusion peptides of most viral fusion-mediating glycoproteins contain a high proportion of hydrophobic amino acids, which facilitates its insertion into target membranes during fusion. In this work we report on the interaction of a 31-residue synthetic peptide (FP31) corresponding to the N terminus of CDV F1 protein with phospholipid membranes composed of various phospholipids, as studied by means of various biophysical techniques. FTIR investigation of FP31 secondary structure in aqueous medium and in membranes resulted in a major proportion of alpha-helical structure which increased upon membrane insertion. Differential scanning calorimetry (DSC) showed that the presence of concentrations of FP31 as low as 0.1 mol%, in mixtures with L-alpha-dimyristoylphosphatidylcholine (DMPC), L-alpha-dipalmitoylphosphatidylcholine (DPPC) and L-alpha-distearoylphosphatidylcholine (DSPC), already affected the thermotropic properties of the gel to liquid-crystalline phase transition. In mixtures with the three lipids, increasing the concentration of peptide made the pretransition to disappear, and lowered and broadened the main transition. This effect was slightly stronger as the acyl chain length of the phospholipid grew larger. In the corresponding partial phase diagrams, no immiscibilities or critical points were observed. FTIR showed that incorporation of 1 mol% of peptide in DPPC shifted the antisymmetric and symmetric CH2 stretching bands to higher values, indicating the existence of an additional disordering of the acyl chain region of the fluid bilayer. FTIR studies of the Cz=O stretching band indicated that incorporation of FP31 into phosphatidylcholine membranes produced a strong dehydration of the polar part of the bilayer. In mixtures with L-alpha-dielaidoylphosphatidylethanolamine (DEPE), increasing FP31 concentrations broadened and shifted to lower temperatures the lamellar to hexagonal-HII phase transition, indicating that this peptide destabilized the bilayer and promoted formation of the hexagonal-HII phase. The results are discussed in terms of lipid-peptide hydrophobic mismatch and its influence on the role of the N-terminal polypeptide of CDV F1 protein in viral membrane fusion.
Aranda
Francisco J
FJ
Departamento de Bioquímica y Biología Molecular-A, Facultad de Veterinaria, Universidad de Murcia, E-30100 Espinardo, Murcia, Spain.
Teruel
José A
JA
Ortiz
Antonio
A
D000818:Animals; D004285:Dogs; D006358:Hot Temperature; D008081:Liposomes / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D013053:Spectrophotometry; D014760:Viral Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism*
D008081:Liposomes; D010455:Peptides; D010743:Phospholipids; D014760:Viral Fusion Proteins; D014764:Viral Proteins; C076619:F protein, Phocine distemper virus