Effect of influenza hemagglutinin fusion peptide on lamellar/inverted phase transitions in dipalmitoleoylphosphatidylethanolamine: implications for membrane fusion mechanisms.
Low mole fractions of viral fusion peptides induce inverted cubic (Q(II)) phases in dipalmitoleoylphosphatidylethanolamine (DiPoPE), a lipid with unsaturated acyl chains that normally forms inverted hexagonal phase (H(II)) above 43 degrees C. The ability to form a Q(II) phase is relevant to the study of membrane fusion: fusion occurs in liposomal systems under conditions where Q(II) phase precursors form, and fusion may be an obligatory step in the lamellar (L(alpha))/Q(II) phase transition. We used X-ray diffraction and time-resolved cryoelectron microscopy (TRC-TEM) to study the effects of the influenza hemagglutinin fusion peptide on the phase behavior and structure of DiPoPE. X-ray diffraction data show that at concentrations of 3-7 mol%, the fusion peptide (FP) induces formation of a Q(II) phase in preference to the H(II) phase. TRC-TEM data show that the FP acts at early stages in the phase transition (i.e. within seconds): at 2-7 mol%, FP decreases or inhibits formation of the L(alpha)/H(II) intermediate morphology observed via TRC-TEM in pure DiPoPE at the same temperature. Our X-ray diffraction data imply that FP either does not affect, or slightly increases, the spontaneous curvature of the host lipid (i.e. either does not affect or tends to destabilize inverted phases, respectively). FP may act in part by affecting the relative stability of two intermediate structures in the phase transition mechanism, as suggested previously. These results indicate a new way in which hydrophobic sequences of membrane proteins may be fusogenic.
Siegel
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DP
Department of Chemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210, USA.
Epand
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RM
D002152:Calorimetry, Differential Scanning; D020285:Cryoelectron Microscopy; D006389:Hemagglutinins, Viral / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D008561:Membrane Fusion; D015394:Molecular Structure; D009975:Orthomyxoviridae; D010714:Phosphatidylethanolamines / Q000737:chemistry*; D013696:Temperature; D014760:Viral Fusion Proteins / Q000737:chemistry*; D014961:X-Ray Diffraction
D006389:Hemagglutinins, Viral; D008081:Liposomes; D010714:Phosphatidylethanolamines; D014760:Viral Fusion Proteins; C060814:hemagglutinin esterase; C007510:1,2-dipalmitoyl-3-phosphatidylethanolamine