title: Fusion peptide-phospholipid noncovalent interactions as observed by nanoelectrospray FTICR-MS.
abstract: Viral infections are propagated by the fusing of the viral membrane with a host cell membrane. Initiation of the fusion process occurs upon perturbation of the membrane of the cell under attack by a subunit of the viral protein known as a fusion peptide. Fusion peptides must insert into the lipid-rich host cell membrane to initiate rupture and merging of the two entities, but much remains unknown about the details of the fusion process. We present detailed electrospray mass spectrometry studies of binding specificities of model fusion peptides P294 and P326 with cell membrane phospholipids, i.e., phosphatidylcholines (PCs, such as 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)) and phosphatidylglycerols (PGs, such as 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DMPG)). The fusion peptides clearly bind more strongly to negatively charged DMPG than to zwitterionic DMPC. Detected binding between P294/P326 and PC/PG in 100% aqueous solution was disrupted by addition of methanol, which is known to weaken hydrophobic interactions; a higher percentage of methanol was needed to destroy a stronger initial binding. Further increases in the methanol volume fraction generally resulted in a reappearance of peptide-lipid binding, with binding strength quotients of 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC)/1,2-dilauroyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DLPG)-peptide complexes rising more steeply than those of DMPC/DMPG-peptide complexes. Compared to fusion peptides P294 and P326, a hydrophilic peptide, fibrinopeptide B, showed much weaker affinity for zwitterionic DMPC, but had moderate binding affinity to negatively charged DMPG in 100% aqueous solutions. However, upon progressive addition of methanol, this hydrophilic peptide showed only a minor initial decrease in binding to DMPG before the detected binding eventually increased. These results contrast with those obtained for the hydrophobic peptides, and offer corroborative evidence that hydrophobic interactions play a key role in the mass spectrometrically observed binding between fusion peptides and phospholipids. Because the rate of viral infection has been found to be pH-dependent, the effect of initial solution pH on peptide-lipid binding was also studied. As the pH was lowered, P326-DMPC binding had a steep and immediate weakening, whereas the P294-DMPC binding was slightly strengthened at pH 3.7 and then gradually weakened with a further decrease in pH. Both P326 and P294 exhibited affinities toward unsaturated lipids; (18:1)PC bound slightly more strongly to P294 than (18:3)PC. These experiments offer further evidence of the ability of electrospray mass spectrometry to provide binding information concerning noncovalent interactions that were established principally by the hydrophobic effect in solution.
authors.affiliation: Department of Chemistry, University of New Orleans, New Orleans, Louisiana 70148, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Frédéric
authors.initials: F
authors.lastname: Heitz
authors.forename: Christian
authors.initials: C
authors.lastname: Le Grimellec
authors.forename: Richard B
authors.initials: RB
authors.lastname: Cole
mesh_terms: D000595:Amino Acid Sequence; D006863:Hydrogen-Ion Concentration; D008561:Membrane Fusion; D008969:Molecular Sequence Data; D036103:Nanotechnology; D010743:Phospholipids / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared; D014760:Viral Fusion Proteins / Q000378:metabolism*
chemical_list: D010743:Phospholipids; D014760:Viral Fusion Proteins