Lipogastrins as potent inhibitors of viral fusion.
The rate and extent of membrane fusion is markedly sensitive to membrane interfacial properties. Lipopeptides with hydrophilic peptide moieties will insert into membranes, leaving the peptide portion at the membrane-water interface. In this work, we have used a lipopeptide composed of the peptide [Nle15]-gastrin-(2-17)-amide covalently linked to 1,2-diacyl-3-mercaptoglycerol-N(alpha)-maleoyl-beta-alanine to give DM-gastrin or DP-gastrin having 14 or 16 carbon atom acyl chains, respectively. The fluorescence emission from the two Trp residues of these lipopeptides exhibited little or no blue shift upon addition of liposomes of egg-phosphatidylethanolamine containing 5 mol% G(D1a). Iodide quenching of DP-gastrin fluorescence was also independent of lipid. These results indicate that the peptide moiety is exposed to the aqueous environment even though the lipopeptide is firmly anchored to the membrane. Both DM and DP-gastrin markedly raise the bilayer to hexagonal phase transition temperature of dipalmitoleoyl phosphatidylethanolamine. However, DM-E5 lowers this phase transition temperature. These lipopeptides have effects on the overall fusion of Sendai virus to liposomes in accord with their opposite effects on lipid curvature. The lipogastrins are potent inhibitors of viral fusion, while DM-E5 slightly promotes this process. Truncated forms of DM-gastrin are also inhibitory to viral fusion, but are less inhibitory than the full lipopeptide. Analysis of the fusion kinetics shows that DP-gastrin causes a reduction in the final extent of fusion and a marked lowering of the fusion rate constant. Binding of Sendai virus to the ganglioside receptor-containing liposomes was not affected. Consideration of the various contributions to the mechanism of inhibition of viral fusion suggests that effects of lipogastrin on membrane intrinsic monolayer curvature is of primary importance.
Epand
R F
RF
Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
Moroder
L
L
Lutz
J
J
Flanagan
T D
TD
Nir
S
S
Epand
R M
RM
D000595:Amino Acid Sequence; D000818:Animals; D002152:Calorimetry, Differential Scanning; D002642:Chick Embryo; D002766:Cholecystokinin / Q000031:analogs & derivatives* / Q000494:pharmacology; D005755:Gastrins / Q000494:pharmacology*; D008561:Membrane Fusion / Q000187:drug effects*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology*; D018108:Respirovirus / Q000187:drug effects / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D014760:Viral Fusion Proteins / Q000037:antagonists & inhibitors*
C079715:DP-gastrin; D005755:Gastrins; D010446:Peptide Fragments; D014760:Viral Fusion Proteins; C079714:DM-gastrin; C083296:dimyristoylmercaptoglycero-N(alpha)maleoyl-beta-alanyl(Thr,Nle)-CCK-9; D002766:Cholecystokinin