Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 2001 Feb;80(2):643–655. doi: 10.1016/S0006-3495(01)76045-6

Implicit solvent model studies of the interactions of the influenza hemagglutinin fusion peptide with lipid bilayers.

D Bechor 1, N Ben-Tal 1
PMCID: PMC1301264  PMID: 11159433

Abstract

The "fusion peptide," a segment of approximately 20 residues of the influenza hemagglutinin (HA), is necessary and sufficient for HA-induced membrane fusion. We used mean-field calculations of the free energy of peptide-membrane association (DeltaG(tot)) to deduce the most probable orientation of the fusion peptide in the membrane. The main contributions to DeltaG(tot) are probably from the electrostatic (DeltaG(el)) and nonpolar (DeltaG(np)) components of the solvation free energy; these were calculated using continuum solvent models. The peptide was described in atomic detail and was modeled as an alpha-helix based on spectroscopic data. The membrane's hydrocarbon region was described as a structureless slab of nonpolar medium embedded in water. All the helix-membrane configurations, which were lower in DeltaG(tot) than the isolated helix in the aqueous phase, were in the same (wide) basin in configurational space. In each, the helix was horizontally adsorbed at the water-bilayer interface with its principal axis parallel to the membrane plane, its hydrophobic face dissolved in the bilayer, and its polar face in the water. The associated DeltaG(tot) value was approximately -8 to -10 kcal/mol (depending on the rotameric state of one of the phenylalanine residues). In contrast, the DeltaG(tot) values associated with experimentally observed oblique orientations were found to be near zero, suggesting they are marginally stable at best. The theoretical model did not take into account the interactions of the polar headgroups with the peptide and peptide-induced membrane deformation effects. Either or both may overcompensate for the DeltaG(tot) difference between the horizontal and oblique orientations.

Full Text

The Full Text of this article is available as a PDF (2.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altmann K. H., Wójcik J., Vásquez M., Scheraga H. A. Helix-coil stability constants for the naturally occurring amino acids in water. XXIII. Proline parameters from random poly (hydroxybutylglutamine-co-L-proline). Biopolymers. 1990;30(1-2):107–120. doi: 10.1002/bip.360300112. [DOI] [PubMed] [Google Scholar]
  2. Ashcroft R. G., Coster H. G., Smith J. R. The molecular organisation of bimolecular lipid membranes. The dielectric structure of the hydrophilic/hydrophobic interface. Biochim Biophys Acta. 1981 Apr 22;643(1):191–204. doi: 10.1016/0005-2736(81)90232-7. [DOI] [PubMed] [Google Scholar]
  3. Ben-Shaul A., Ben-Tal N., Honig B. Statistical thermodynamic analysis of peptide and protein insertion into lipid membranes. Biophys J. 1996 Jul;71(1):130–137. doi: 10.1016/S0006-3495(96)79208-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ben-Tal N., Ben-Shaul A., Nicholls A., Honig B. Free-energy determinants of alpha-helix insertion into lipid bilayers. Biophys J. 1996 Apr;70(4):1803–1812. doi: 10.1016/S0006-3495(96)79744-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ben-Tal N., Honig B., Bagdassarian C. K., Ben-Shaul A. Association entropy in adsorption processes. Biophys J. 2000 Sep;79(3):1180–1187. doi: 10.1016/S0006-3495(00)76372-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ben-Tal N., Honig B., Miller C., McLaughlin S. Electrostatic binding of proteins to membranes. Theoretical predictions and experimental results with charybdotoxin and phospholipid vesicles. Biophys J. 1997 Oct;73(4):1717–1727. doi: 10.1016/S0006-3495(97)78203-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ben-Tal N., Honig B., Peitzsch R. M., Denisov G., McLaughlin S. Binding of small basic peptides to membranes containing acidic lipids: theoretical models and experimental results. Biophys J. 1996 Aug;71(2):561–575. doi: 10.1016/S0006-3495(96)79280-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ben-Tal N., Sitkoff D., Bransburg-Zabary S., Nachliel E., Gutman M. Theoretical calculations of the permeability of monensin-cation complexes in model bio-membranes. Biochim Biophys Acta. 2000 Jun 1;1466(1-2):221–233. doi: 10.1016/s0005-2736(00)00156-5. [DOI] [PubMed] [Google Scholar]
  9. Bernèche S., Nina M., Roux B. Molecular dynamics simulation of melittin in a dimyristoylphosphatidylcholine bilayer membrane. Biophys J. 1998 Oct;75(4):1603–1618. doi: 10.1016/S0006-3495(98)77604-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Biggin P. C., Breed J., Son H. S., Sansom M. S. Simulation studies of alamethicin-bilayer interactions. Biophys J. 1997 Feb;72(2 Pt 1):627–636. doi: 10.1016/s0006-3495(97)78701-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Brasseur R., Vandenbranden M., Cornet B., Burny A., Ruysschaert J. M. Orientation into the lipid bilayer of an asymmetric amphipathic helical peptide located at the N-terminus of viral fusion proteins. Biochim Biophys Acta. 1990 Nov 16;1029(2):267–273. doi: 10.1016/0005-2736(90)90163-i. [DOI] [PubMed] [Google Scholar]
  12. Bullough P. A., Hughson F. M., Skehel J. J., Wiley D. C. Structure of influenza haemagglutinin at the pH of membrane fusion. Nature. 1994 Sep 1;371(6492):37–43. doi: 10.1038/371037a0. [DOI] [PubMed] [Google Scholar]
  13. Buser C. A., Sigal C. T., Resh M. D., McLaughlin S. Membrane binding of myristylated peptides corresponding to the NH2 terminus of Src. Biochemistry. 1994 Nov 8;33(44):13093–13101. doi: 10.1021/bi00248a019. [DOI] [PubMed] [Google Scholar]
  14. Carr C. M., Kim P. S. A spring-loaded mechanism for the conformational change of influenza hemagglutinin. Cell. 1993 May 21;73(4):823–832. doi: 10.1016/0092-8674(93)90260-w. [DOI] [PubMed] [Google Scholar]
  15. Carr C. M., Kim P. S. Flu virus invasion: halfway there. Science. 1994 Oct 14;266(5183):234–236. doi: 10.1126/science.7939658. [DOI] [PubMed] [Google Scholar]
  16. Chen J., Lee K. H., Steinhauer D. A., Stevens D. J., Skehel J. J., Wiley D. C. Structure of the hemagglutinin precursor cleavage site, a determinant of influenza pathogenicity and the origin of the labile conformation. Cell. 1998 Oct 30;95(3):409–417. doi: 10.1016/s0092-8674(00)81771-7. [DOI] [PubMed] [Google Scholar]
  17. Chernomordik L. V., Leikina E., Frolov V., Bronk P., Zimmerberg J. An early stage of membrane fusion mediated by the low pH conformation of influenza hemagglutinin depends upon membrane lipids. J Cell Biol. 1997 Jan 13;136(1):81–93. doi: 10.1083/jcb.136.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Colotto A., Epand R. M. Structural study of the relationship between the rate of membrane fusion and the ability of the fusion peptide of influenza virus to perturb bilayers. Biochemistry. 1997 Jun 24;36(25):7644–7651. doi: 10.1021/bi970382u. [DOI] [PubMed] [Google Scholar]
  19. Dilger J. P., Benz R. Optical and electrical properties of thin monoolein lipid bilayers. J Membr Biol. 1985;85(2):181–189. doi: 10.1007/BF01871270. [DOI] [PubMed] [Google Scholar]
  20. Durell S. R., Martin I., Ruysschaert J. M., Shai Y., Blumenthal R. What studies of fusion peptides tell us about viral envelope glycoprotein-mediated membrane fusion (review). Mol Membr Biol. 1997 Jul-Sep;14(3):97–112. doi: 10.3109/09687689709048170. [DOI] [PubMed] [Google Scholar]
  21. Efremov R. G., Nolde D. E., Vergoten G., Arseniev A. S. A solvent model for simulations of peptides in bilayers. I. Membrane-promoting alpha-helix formation. Biophys J. 1999 May;76(5):2448–2459. doi: 10.1016/S0006-3495(99)77400-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Efremov R. G., Nolde D. E., Volynsky P. E., Chernyavsky A. A., Dubovskii P. V., Arseniev A. S. Factors important for fusogenic activity of peptides: molecular modeling study of analogs of fusion peptide of influenza virus hemagglutinin. FEBS Lett. 1999 Nov 26;462(1-2):205–210. doi: 10.1016/s0014-5793(99)01505-7. [DOI] [PubMed] [Google Scholar]
  23. Engelman D. M., Steitz T. A. The spontaneous insertion of proteins into and across membranes: the helical hairpin hypothesis. Cell. 1981 Feb;23(2):411–422. doi: 10.1016/0092-8674(81)90136-7. [DOI] [PubMed] [Google Scholar]
  24. Gething M. J., Doms R. W., York D., White J. Studies on the mechanism of membrane fusion: site-specific mutagenesis of the hemagglutinin of influenza virus. J Cell Biol. 1986 Jan;102(1):11–23. doi: 10.1083/jcb.102.1.11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Gilson M. K. Theory of electrostatic interactions in macromolecules. Curr Opin Struct Biol. 1995 Apr;5(2):216–223. doi: 10.1016/0959-440x(95)80079-4. [DOI] [PubMed] [Google Scholar]
  26. Gray C., Tatulian S. A., Wharton S. A., Tamm L. K. Effect of the N-terminal glycine on the secondary structure, orientation, and interaction of the influenza hemagglutinin fusion peptide with lipid bilayers. Biophys J. 1996 May;70(5):2275–2286. doi: 10.1016/S0006-3495(96)79793-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Han X., Steinhauer D. A., Wharton S. A., Tamm L. K. Interaction of mutant influenza virus hemagglutinin fusion peptides with lipid bilayers: probing the role of hydrophobic residue size in the central region of the fusion peptide. Biochemistry. 1999 Nov 9;38(45):15052–15059. doi: 10.1021/bi991232h. [DOI] [PubMed] [Google Scholar]
  28. Harter C., James P., Bächi T., Semenza G., Brunner J. Hydrophobic binding of the ectodomain of influenza hemagglutinin to membranes occurs through the "fusion peptide". J Biol Chem. 1989 Apr 15;264(11):6459–6464. [PubMed] [Google Scholar]
  29. Hernandez L. D., Hoffman L. R., Wolfsberg T. G., White J. M. Virus-cell and cell-cell fusion. Annu Rev Cell Dev Biol. 1996;12:627–661. doi: 10.1146/annurev.cellbio.12.1.627. [DOI] [PubMed] [Google Scholar]
  30. Honig B., Nicholls A. Classical electrostatics in biology and chemistry. Science. 1995 May 26;268(5214):1144–1149. doi: 10.1126/science.7761829. [DOI] [PubMed] [Google Scholar]
  31. Jacobs R. E., White S. H. The nature of the hydrophobic binding of small peptides at the bilayer interface: implications for the insertion of transbilayer helices. Biochemistry. 1989 Apr 18;28(8):3421–3437. doi: 10.1021/bi00434a042. [DOI] [PubMed] [Google Scholar]
  32. Jähnig F. Thermodynamics and kinetics of protein incorporation into membranes. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3691–3695. doi: 10.1073/pnas.80.12.3691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kessel A., Cafiso D. S., Ben-Tal N. Continuum solvent model calculations of alamethicin-membrane interactions: thermodynamic aspects. Biophys J. 2000 Feb;78(2):571–583. doi: 10.1016/S0006-3495(00)76617-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Kessel A., Schulten K., Ben-Tal N. Calculations suggest a pathway for the transverse diffusion of a hydrophobic peptide across a lipid bilayer. Biophys J. 2000 Nov;79(5):2322–2330. doi: 10.1016/S0006-3495(00)76478-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Kurumbail R. G., Stevens A. M., Gierse J. K., McDonald J. J., Stegeman R. A., Pak J. Y., Gildehaus D., Miyashiro J. M., Penning T. D., Seibert K. Structural basis for selective inhibition of cyclooxygenase-2 by anti-inflammatory agents. Nature. 1996 Dec 19;384(6610):644–648. doi: 10.1038/384644a0. [DOI] [PubMed] [Google Scholar]
  36. Lear J. D., DeGrado W. F. Membrane binding and conformational properties of peptides representing the NH2 terminus of influenza HA-2. J Biol Chem. 1987 May 15;262(14):6500–6505. [PubMed] [Google Scholar]
  37. Lüneberg J., Martin I., Nüssler F., Ruysschaert J. M., Herrmann A. Structure and topology of the influenza virus fusion peptide in lipid bilayers. J Biol Chem. 1995 Nov 17;270(46):27606–27614. doi: 10.1074/jbc.270.46.27606. [DOI] [PubMed] [Google Scholar]
  38. Macosko J. C., Kim C. H., Shin Y. K. The membrane topology of the fusion peptide region of influenza hemagglutinin determined by spin-labeling EPR. J Mol Biol. 1997 Apr 18;267(5):1139–1148. doi: 10.1006/jmbi.1997.0931. [DOI] [PubMed] [Google Scholar]
  39. Milik M., Skolnick J. Insertion of peptide chains into lipid membranes: an off-lattice Monte Carlo dynamics model. Proteins. 1993 Jan;15(1):10–25. doi: 10.1002/prot.340150104. [DOI] [PubMed] [Google Scholar]
  40. Moll T. S., Thompson T. E. Semisynthetic proteins: model systems for the study of the insertion of hydrophobic peptides into preformed lipid bilayers. Biochemistry. 1994 Dec 27;33(51):15469–15482. doi: 10.1021/bi00255a029. [DOI] [PubMed] [Google Scholar]
  41. Nakamura H. Roles of electrostatic interaction in proteins. Q Rev Biophys. 1996 Feb;29(1):1–90. doi: 10.1017/s0033583500005746. [DOI] [PubMed] [Google Scholar]
  42. Nicholls A., Sharp K. A., Honig B. Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins. 1991;11(4):281–296. doi: 10.1002/prot.340110407. [DOI] [PubMed] [Google Scholar]
  43. Nozaki Y., Tanford C. The solubility of amino acids and two glycine peptides in aqueous ethanol and dioxane solutions. Establishment of a hydrophobicity scale. J Biol Chem. 1971 Apr 10;246(7):2211–2217. [PubMed] [Google Scholar]
  44. Picot D., Loll P. J., Garavito R. M. The X-ray crystal structure of the membrane protein prostaglandin H2 synthase-1. Nature. 1994 Jan 20;367(6460):243–249. doi: 10.1038/367243a0. [DOI] [PubMed] [Google Scholar]
  45. Sharp K. A., Nicholls A., Fine R. F., Honig B. Reconciling the magnitude of the microscopic and macroscopic hydrophobic effects. Science. 1991 Apr 5;252(5002):106–109. doi: 10.1126/science.2011744. [DOI] [PubMed] [Google Scholar]
  46. Siegel D. P., Epand R. M. Effect of influenza hemagglutinin fusion peptide on lamellar/inverted phase transitions in dipalmitoleoylphosphatidylethanolamine: implications for membrane fusion mechanisms. Biochim Biophys Acta. 2000 Sep 29;1468(1-2):87–98. doi: 10.1016/s0005-2736(00)00246-7. [DOI] [PubMed] [Google Scholar]
  47. Steinhauer D. A., Wharton S. A., Skehel J. J., Wiley D. C. Studies of the membrane fusion activities of fusion peptide mutants of influenza virus hemagglutinin. J Virol. 1995 Nov;69(11):6643–6651. doi: 10.1128/jvi.69.11.6643-6651.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Takahashi S. Conformation of membrane fusion-active 20-residue peptides with or without lipid bilayers. Implication of alpha-helix formation for membrane fusion. Biochemistry. 1990 Jul 3;29(26):6257–6264. doi: 10.1021/bi00478a021. [DOI] [PubMed] [Google Scholar]
  49. Thorgeirsson T. E., Russell C. J., King D. S., Shin Y. K. Direct determination of the membrane affinities of individual amino acids. Biochemistry. 1996 Feb 13;35(6):1803–1809. doi: 10.1021/bi952300c. [DOI] [PubMed] [Google Scholar]
  50. Warshel A., Papazyan A. Electrostatic effects in macromolecules: fundamental concepts and practical modeling. Curr Opin Struct Biol. 1998 Apr;8(2):211–217. doi: 10.1016/s0959-440x(98)80041-9. [DOI] [PubMed] [Google Scholar]
  51. White J. M. Membrane fusion. Science. 1992 Nov 6;258(5084):917–924. doi: 10.1126/science.1439803. [DOI] [PubMed] [Google Scholar]
  52. White S. H., Wimley W. C. Membrane protein folding and stability: physical principles. Annu Rev Biophys Biomol Struct. 1999;28:319–365. doi: 10.1146/annurev.biophys.28.1.319. [DOI] [PubMed] [Google Scholar]
  53. Wilson I. A., Skehel J. J., Wiley D. C. Structure of the haemagglutinin membrane glycoprotein of influenza virus at 3 A resolution. Nature. 1981 Jan 29;289(5796):366–373. doi: 10.1038/289366a0. [DOI] [PubMed] [Google Scholar]
  54. Yang A. S., Honig B. Free energy determinants of secondary structure formation: I. alpha-Helices. J Mol Biol. 1995 Sep 22;252(3):351–365. doi: 10.1006/jmbi.1995.0502. [DOI] [PubMed] [Google Scholar]
  55. Zhou Z., Macosko J. C., Hughes D. W., Sayer B. G., Hawes J., Epand R. M. 15N NMR study of the ionization properties of the influenza virus fusion peptide in zwitterionic phospholipid dispersions. Biophys J. 2000 May;78(5):2418–2425. doi: 10.1016/S0006-3495(00)76785-3. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES