Skip to main content
Biophysical Journal logoLink to Biophysical Journal
. 1995 Sep;69(3):922–929. doi: 10.1016/S0006-3495(95)79966-0

The hemifusion intermediate and its conversion to complete fusion: regulation by membrane composition.

L Chernomordik 1, A Chanturiya 1, J Green 1, J Zimmerberg 1
PMCID: PMC1236321  PMID: 8519992

Abstract

To fuse, membranes must bend. The energy of each lipid monolayer with respect to bending is minimized at the spontaneous curvature of the monolayer. Two lipids known to promote opposite spontaneous curvatures, lysophosphatidylcholine and arachidonic acid, were added to different sides of planar phospholipid membranes. Lysophosphatidylcholine added to the contacting monolayers of fusing membranes inhibited the hemifusion we observed between lipid vesicles and planar membranes. In contrast, fusion pore formation depended upon the distal monolayer of the planar membrane; lysophosphatidylcholine promoted and arachidonic acid inhibited. Thus, the intermediates of hemifusion and fusion pores in phospholipid membranes involve different membrane monolayers and may have opposite net curvatures, Biological fusion may proceed through similar intermediates.

Full text

PDF
922

Images in this article

Selected References

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

  1. Armstrong S. K., Parr T. R., Jr, Parker C. D., Hancock R. E. Bordetella pertussis major outer membrane porin protein forms small, anion-selective channels in lipid bilayer membranes. J Bacteriol. 1986 Apr;166(1):212–216. doi: 10.1128/jb.166.1.212-216.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bentz J., Ellens H., Lai M. Z., Szoka F. C., Jr On the correlation between HII phase and the contact-induced destabilization of phosphatidylethanolamine-containing membranes. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5742–5745. doi: 10.1073/pnas.82.17.5742. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chernomordik L. V., Melikyan G. B., Chizmadzhev Y. A. Biomembrane fusion: a new concept derived from model studies using two interacting planar lipid bilayers. Biochim Biophys Acta. 1987 Oct 5;906(3):309–352. doi: 10.1016/0304-4157(87)90016-5. [DOI] [PubMed] [Google Scholar]
  4. Chernomordik L. V., Vogel S. S., Sokoloff A., Onaran H. O., Leikina E. A., Zimmerberg J. Lysolipids reversibly inhibit Ca(2+)-, GTP- and pH-dependent fusion of biological membranes. FEBS Lett. 1993 Feb 22;318(1):71–76. doi: 10.1016/0014-5793(93)81330-3. [DOI] [PubMed] [Google Scholar]
  5. Chernomordik L., Leikina E., Cho M. S., Zimmerberg J. Control of baculovirus gp64-induced syncytium formation by membrane lipid composition. J Virol. 1995 May;69(5):3049–3058. doi: 10.1128/jvi.69.5.3049-3058.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chung H., Caffrey M. The curvature elastic-energy function of the lipid-water cubic mesophase. Nature. 1994 Mar 17;368(6468):224–226. doi: 10.1038/368224a0. [DOI] [PubMed] [Google Scholar]
  7. Cohen F. S., Akabas M. H., Finkelstein A. Osmotic swelling of phospholipid vesicles causes them to fuse with a planar phospholipid bilayer membrane. Science. 1982 Jul 30;217(4558):458–460. doi: 10.1126/science.6283637. [DOI] [PubMed] [Google Scholar]
  8. Cohen F. S., Akabas M. H., Zimmerberg J., Finkelstein A. Parameters affecting the fusion of unilamellar phospholipid vesicles with planar bilayer membranes. J Cell Biol. 1984 Mar;98(3):1054–1062. doi: 10.1083/jcb.98.3.1054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cohen F. S., Niles W. D. Reconstituting channels into planar membranes: a conceptual framework and methods for fusing vesicles to planar bilayer phospholipid membranes. Methods Enzymol. 1993;220:50–68. doi: 10.1016/0076-6879(93)20073-c. [DOI] [PubMed] [Google Scholar]
  10. Cohen F. S., Zimmerberg J., Finkelstein A. Fusion of phospholipid vesicles with planar phospholipid bilayer membranes. II. Incorporation of a vesicular membrane marker into the planar membrane. J Gen Physiol. 1980 Mar;75(3):251–270. doi: 10.1085/jgp.75.3.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Düzgüneş N., Straubinger R. M., Baldwin P. A., Friend D. S., Papahadjopoulos D. Proton-induced fusion of oleic acid-phosphatidylethanolamine liposomes. Biochemistry. 1985 Jun 18;24(13):3091–3098. doi: 10.1021/bi00334a004. [DOI] [PubMed] [Google Scholar]
  12. Elhai J., Scandella C. J. Arachidonic acid and other fatty acids inhibit secretion from sea urchin eggs. Exp Cell Res. 1983 Oct;148(1):63–71. doi: 10.1016/0014-4827(83)90187-8. [DOI] [PubMed] [Google Scholar]
  13. Ellens H., Bentz J., Szoka F. C. Destabilization of phosphatidylethanolamine liposomes at the hexagonal phase transition temperature. Biochemistry. 1986 Jan 28;25(2):285–294. doi: 10.1021/bi00350a001. [DOI] [PubMed] [Google Scholar]
  14. Epand R. M. Diacylglycerols, lysolecithin, or hydrocarbons markedly alter the bilayer to hexagonal phase transition temperature of phosphatidylethanolamines. Biochemistry. 1985 Dec 3;24(25):7092–7095. doi: 10.1021/bi00346a011. [DOI] [PubMed] [Google Scholar]
  15. Epand R. M., Epand R. F., Ahmed N., Chen R. Promotion of hexagonal phase formation and lipid mixing by fatty acids with varying degrees of unsaturation. Chem Phys Lipids. 1991 Jan-Feb;57(1):75–80. doi: 10.1016/0009-3084(91)90051-c. [DOI] [PubMed] [Google Scholar]
  16. Helfrich W. Elastic properties of lipid bilayers: theory and possible experiments. Z Naturforsch C. 1973 Nov-Dec;28(11):693–703. doi: 10.1515/znc-1973-11-1209. [DOI] [PubMed] [Google Scholar]
  17. Hope M. J., Cullis P. R. The role of nonbilayer lipid structures in the fusion of human erythrocytes induced by lipid fusogens. Biochim Biophys Acta. 1981 Jan 8;640(1):82–90. doi: 10.1016/0005-2736(81)90533-2. [DOI] [PubMed] [Google Scholar]
  18. Hui S. W., Sen A. Effects of lipid packing on polymorphic phase behavior and membrane properties. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5825–5829. doi: 10.1073/pnas.86.15.5825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hui S. W., Stewart T. P., Boni L. T., Yeagle P. L. Membrane fusion through point defects in bilayers. Science. 1981 May 22;212(4497):921–923. doi: 10.1126/science.7233185. [DOI] [PubMed] [Google Scholar]
  20. Kemble G. W., Danieli T., White J. M. Lipid-anchored influenza hemagglutinin promotes hemifusion, not complete fusion. Cell. 1994 Jan 28;76(2):383–391. doi: 10.1016/0092-8674(94)90344-1. [DOI] [PubMed] [Google Scholar]
  21. Kozlov M. M., Leikin S. L., Chernomordik L. V., Markin V. S., Chizmadzhev Y. A. Stalk mechanism of vesicle fusion. Intermixing of aqueous contents. Eur Biophys J. 1989;17(3):121–129. doi: 10.1007/BF00254765. [DOI] [PubMed] [Google Scholar]
  22. Kozlov M. M., Leikin S., Rand R. P. Bending, hydration and interstitial energies quantitatively account for the hexagonal-lamellar-hexagonal reentrant phase transition in dioleoylphosphatidylethanolamine. Biophys J. 1994 Oct;67(4):1603–1611. doi: 10.1016/S0006-3495(94)80633-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kumar V. V., Malewicz B., Baumann W. J. Lysophosphatidylcholine stabilizes small unilamellar phosphatidylcholine vesicles. Phosphorus-31 NMR evidence for the "wedge" effect. Biophys J. 1989 Apr;55(4):789–792. doi: 10.1016/S0006-3495(89)82877-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Leikin S. L., Kozlov M. M., Chernomordik L. V., Markin V. S., Chizmadzhev Y. A. Membrane fusion: overcoming of the hydration barrier and local restructuring. J Theor Biol. 1987 Dec 21;129(4):411–425. doi: 10.1016/s0022-5193(87)80021-8. [DOI] [PubMed] [Google Scholar]
  25. Luzzati V., Gulik-Krzywicki T., Rivas E., Reiss-Husson F., Rand R. P. X-ray study of model systems: structure of the lipid-water phases in correlation with the chemical composition of the lipids. J Gen Physiol. 1968 May 1;51(5):37–43. [PMC free article] [PubMed] [Google Scholar]
  26. MacDonald R. C., MacDonald R. I., Menco B. P., Takeshita K., Subbarao N. K., Hu L. R. Small-volume extrusion apparatus for preparation of large, unilamellar vesicles. Biochim Biophys Acta. 1991 Jan 30;1061(2):297–303. doi: 10.1016/0005-2736(91)90295-j. [DOI] [PubMed] [Google Scholar]
  27. MacDonald R. I. Characteristics of self-quenching of the fluorescence of lipid-conjugated rhodamine in membranes. J Biol Chem. 1990 Aug 15;265(23):13533–13539. [PubMed] [Google Scholar]
  28. Niles W. D., Cohen F. S. Video fluorescence microscopy studies of phospholipid vesicle fusion with a planar phospholipid membrane. Nature of membrane-membrane interactions and detection of release of contents. J Gen Physiol. 1987 Nov;90(5):703–735. doi: 10.1085/jgp.90.5.703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Niles W. D., Levis R. A., Cohen F. S. Planar bilayer membranes made from phospholipid monolayers form by a thinning process. Biophys J. 1988 Mar;53(3):327–335. doi: 10.1016/S0006-3495(88)83110-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ornberg R. L., Reese T. S. Beginning of exocytosis captured by rapid-freezing of Limulus amebocytes. J Cell Biol. 1981 Jul;90(1):40–54. doi: 10.1083/jcb.90.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Perin M. S., MacDonald R. C. Interactions of liposomes with planar bilayer membranes. J Membr Biol. 1989 Aug;109(3):221–232. doi: 10.1007/BF01870279. [DOI] [PubMed] [Google Scholar]
  32. Pinto da Silva P., Nogueira M. L. Membrane fusion during secretion. A hypothesis based on electron microscope observation of Phytophthora Palmivora zoospores during encystment. J Cell Biol. 1977 Apr;73(1):161–181. doi: 10.1083/jcb.73.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rand R. P., Fuller N. L., Gruner S. M., Parsegian V. A. Membrane curvature, lipid segregation, and structural transitions for phospholipids under dual-solvent stress. Biochemistry. 1990 Jan 9;29(1):76–87. doi: 10.1021/bi00453a010. [DOI] [PubMed] [Google Scholar]
  34. Siegel D. P. Energetics of intermediates in membrane fusion: comparison of stalk and inverted micellar intermediate mechanisms. Biophys J. 1993 Nov;65(5):2124–2140. doi: 10.1016/S0006-3495(93)81256-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Song L. Y., Ahkong Q. F., Georgescauld D., Lucy J. A. Membrane fusion without cytoplasmic fusion (hemi-fusion) in erythrocytes that are subjected to electrical breakdown. Biochim Biophys Acta. 1991 May 31;1065(1):54–62. doi: 10.1016/0005-2736(91)90010-6. [DOI] [PubMed] [Google Scholar]
  36. Stamatatos L., Düzgüneş N. Simian immunodeficiency virus (SIVmac251) membrane lipid mixing with human CD4+ and CD4- cell lines in vitro does not necessarily result in internalization of the viral core proteins and productive infection. J Gen Virol. 1993 Jun;74(Pt 6):1043–1054. doi: 10.1099/0022-1317-74-6-1043. [DOI] [PubMed] [Google Scholar]
  37. Tate M. W., Eikenberry E. F., Turner D. C., Shyamsunder E., Gruner S. M. Nonbilayer phases of membrane lipids. Chem Phys Lipids. 1991 Mar;57(2-3):147–164. doi: 10.1016/0009-3084(91)90073-k. [DOI] [PubMed] [Google Scholar]
  38. Tse F. W., Iwata A., Almers W. Membrane flux through the pore formed by a fusogenic viral envelope protein during cell fusion. J Cell Biol. 1993 May;121(3):543–552. doi: 10.1083/jcb.121.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Vogel S. S., Leikina E. A., Chernomordik L. V. Lysophosphatidylcholine reversibly arrests exocytosis and viral fusion at a stage between triggering and membrane merger. J Biol Chem. 1993 Dec 5;268(34):25764–25768. [PubMed] [Google Scholar]
  40. Walter A., Yeagle P. L., Siegel D. P. Diacylglycerol and hexadecane increase divalent cation-induced lipid mixing rates between phosphatidylserine large unilamellar vesicles. Biophys J. 1994 Feb;66(2 Pt 1):366–376. doi: 10.1016/s0006-3495(94)80786-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Weltzien H. U. Cytolytic and membrane-perturbing properties of lysophosphatidylcholine. Biochim Biophys Acta. 1979 Aug 20;559(2-3):259–287. doi: 10.1016/0304-4157(79)90004-2. [DOI] [PubMed] [Google Scholar]
  42. Woodbury D. J., Hall J. E. Role of channels in the fusion of vesicles with a planar bilayer. Biophys J. 1988 Dec;54(6):1053–1063. doi: 10.1016/S0006-3495(88)83042-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Zimmerberg J., Blumenthal R., Sarkar D. P., Curran M., Morris S. J. Restricted movement of lipid and aqueous dyes through pores formed by influenza hemagglutinin during cell fusion. J Cell Biol. 1994 Dec;127(6 Pt 2):1885–1894. doi: 10.1083/jcb.127.6.1885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zimmerberg J., Cohen F. S., Finkelstein A. Fusion of phospholipid vesicles with planar phospholipid bilayer membranes. I. Discharge of vesicular contents across the planar membrane. J Gen Physiol. 1980 Mar;75(3):241–250. doi: 10.1085/jgp.75.3.241. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES