Skip to main content
PMC home page
Biophysical Journal logoLink to Biophysical Journal
. 2001 Feb;80(2):801–811. doi: 10.1016/S0006-3495(01)76059-6

Structure, location, and lipid perturbations of melittin at the membrane interface.

K Hristova 1, C E Dempsey 1, S H White 1
PMCID: PMC1301278  PMID: 11159447

Abstract

Melittin is arguably the most widely studied amphipathic, membrane-lytic alpha-helical peptide. Although several lines of evidence suggest an interfacial membrane location at low concentrations, melittin's exact position and depth of penetration into the hydrocarbon core are unknown. Furthermore, the structural basis for its lytic action remains largely a matter of conjecture. Using a novel x-ray absolute-scale refinement method, we have now determined the location, orientation, and likely conformation of monomeric melittin in oriented phosphocholine lipid multilayers. Its helical axis is aligned parallel to the bilayer plane at the depth of the glycerol groups, but its average conformation differs from the crystallographic structure. As observed earlier for another amphipathic alpha-helical peptide, the lipid perturbations induced by melittin are remarkably modest. Small bilayer perturbations thus appear to be a general feature of amphipathic helices at low concentrations. In contrast, a dimeric form of melittin causes larger structural perturbations under otherwise identical conditions. These results provide direct structural evidence that self-association of amphipathic helices may be the crucial initial step toward membrane lysis.

Full Text

The Full Text of this article is available as a PDF (836.6 KB).

Selected References

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

  1. Altenbach C., Froncisz W., Hyde J. S., Hubbell W. L. Conformation of spin-labeled melittin at membrane surfaces investigated by pulse saturation recovery and continuous wave power saturation electron paramagnetic resonance. Biophys J. 1989 Dec;56(6):1183–1191. doi: 10.1016/S0006-3495(89)82765-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Altenbach C., Hubbell W. L. The aggregation state of spin-labeled melittin in solution and bound to phospholipid membranes: evidence that membrane-bound melittin is monomeric. Proteins. 1988;3(4):230–242. doi: 10.1002/prot.340030404. [DOI] [PubMed] [Google Scholar]
  3. Bachar M., Becker O. M. Protein-induced membrane disorder: a molecular dynamics study of melittin in a dipalmitoylphosphatidylcholine bilayer. Biophys J. 2000 Mar;78(3):1359–1375. doi: 10.1016/S0006-3495(00)76690-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baumann G., Mueller P. A molecular model of membrane excitability. J Supramol Struct. 1974;2(5-6):538–557. doi: 10.1002/jss.400020504. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Bradshaw J. P., Dempsey C. E., Watts A. A combined X-ray and neutron diffraction study of selectively deuterated melittin in phospholipid bilayers: effect of pH. Mol Membr Biol. 1994 Apr-Jun;11(2):79–86. doi: 10.3109/09687689409162224. [DOI] [PubMed] [Google Scholar]
  7. DeGrado W. F., Musso G. F., Lieber M., Kaiser E. T., Kézdy F. J. Kinetics and mechanism of hemolysis induced by melittin and by a synthetic melittin analogue. Biophys J. 1982 Jan;37(1):329–338. doi: 10.1016/S0006-3495(82)84681-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dempsey C. E., Butler G. S. Helical structure and orientation of melittin in dispersed phospholipid membranes from amide exchange analysis in situ. Biochemistry. 1992 Dec 8;31(48):11973–11977. doi: 10.1021/bi00163a003. [DOI] [PubMed] [Google Scholar]
  9. Dempsey C. E. The actions of melittin on membranes. Biochim Biophys Acta. 1990 May 7;1031(2):143–161. doi: 10.1016/0304-4157(90)90006-x. [DOI] [PubMed] [Google Scholar]
  10. Fox R. O., Jr, Richards F. M. A voltage-gated ion channel model inferred from the crystal structure of alamethicin at 1.5-A resolution. Nature. 1982 Nov 25;300(5890):325–330. doi: 10.1038/300325a0. [DOI] [PubMed] [Google Scholar]
  11. Franks N. P., Arunachalam T., Caspi E. A direct method for determination of membrane electron density profiles on an absolute scale. Nature. 1978 Nov 30;276(5687):530–532. doi: 10.1038/276530a0. [DOI] [PubMed] [Google Scholar]
  12. Franks N. P., Levine Y. K. Low-angle x-ray diffraction. Mol Biol Biochem Biophys. 1981;31:437–487. doi: 10.1007/978-3-642-81537-9_9. [DOI] [PubMed] [Google Scholar]
  13. Frey S., Tamm L. K. Orientation of melittin in phospholipid bilayers. A polarized attenuated total reflection infrared study. Biophys J. 1991 Oct;60(4):922–930. doi: 10.1016/S0006-3495(91)82126-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Habermann E. Bee and wasp venoms. Science. 1972 Jul 28;177(4046):314–322. doi: 10.1126/science.177.4046.314. [DOI] [PubMed] [Google Scholar]
  15. Hall J. E., Vodyanoy I., Balasubramanian T. M., Marshall G. R. Alamethicin. A rich model for channel behavior. Biophys J. 1984 Jan;45(1):233–247. doi: 10.1016/S0006-3495(84)84151-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hristova K., White S. H. Determination of the hydrocarbon core structure of fluid dioleoylphosphocholine (DOPC) bilayers by x-ray diffraction using specific bromination of the double-bonds: effect of hydration. Biophys J. 1998 May;74(5):2419–2433. doi: 10.1016/S0006-3495(98)77950-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hristova K., Wimley W. C., Mishra V. K., Anantharamiah G. M., Segrest J. P., White S. H. An amphipathic alpha-helix at a membrane interface: a structural study using a novel X-ray diffraction method. J Mol Biol. 1999 Jul 2;290(1):99–117. doi: 10.1006/jmbi.1999.2840. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. John E., Jähnig F. Aggregation state of melittin in lipid vesicle membranes. Biophys J. 1991 Aug;60(2):319–328. doi: 10.1016/S0006-3495(91)82056-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Katsu T., Ninomiya C., Kuroko M., Kobayashi H., Hirota T., Fujita Y. Action mechanism of amphipathic peptides gramicidin S and melittin on erythrocyte membrane. Biochim Biophys Acta. 1988 Mar 22;939(1):57–63. doi: 10.1016/0005-2736(88)90047-8. [DOI] [PubMed] [Google Scholar]
  21. Ladokhin A. S., Selsted M. E., White S. H. Bilayer interactions of indolicidin, a small antimicrobial peptide rich in tryptophan, proline, and basic amino acids. Biophys J. 1997 Feb;72(2 Pt 1):794–805. doi: 10.1016/s0006-3495(97)78713-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ladokhin A. S., Selsted M. E., White S. H. CD spectra of indolicidin antimicrobial peptides suggest turns, not polyproline helix. Biochemistry. 1999 Sep 21;38(38):12313–12319. doi: 10.1021/bi9907936. [DOI] [PubMed] [Google Scholar]
  23. Ladokhin A. S., Selsted M. E., White S. H. Sizing membrane pores in lipid vesicles by leakage of co-encapsulated markers: pore formation by melittin. Biophys J. 1997 Apr;72(4):1762–1766. doi: 10.1016/S0006-3495(97)78822-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ladokhin A. S., White S. H. Folding of amphipathic alpha-helices on membranes: energetics of helix formation by melittin. J Mol Biol. 1999 Jan 29;285(4):1363–1369. doi: 10.1006/jmbi.1998.2346. [DOI] [PubMed] [Google Scholar]
  25. MacNaughtan W., Snook K. A., Caspi E., Franks N. P. An X-ray diffraction analysis of oriented lipid multilayers containing basic proteins. Biochim Biophys Acta. 1985 Aug 27;818(2):132–148. doi: 10.1016/0005-2736(85)90556-5. [DOI] [PubMed] [Google Scholar]
  26. Makhatadze G. I., Medvedkin V. N., Privalov P. L. Partial molar volumes of polypeptides and their constituent groups in aqueous solution over a broad temperature range. Biopolymers. 1990;30(11-12):1001–1010. doi: 10.1002/bip.360301102. [DOI] [PubMed] [Google Scholar]
  27. Matsuzaki K., Yoneyama S., Miyajima K. Pore formation and translocation of melittin. Biophys J. 1997 Aug;73(2):831–838. doi: 10.1016/S0006-3495(97)78115-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mishra V. K., Palgunachari M. N., Segrest J. P., Anantharamaiah G. M. Interactions of synthetic peptide analogs of the class A amphipathic helix with lipids. Evidence for the snorkel hypothesis. J Biol Chem. 1994 Mar 11;269(10):7185–7191. [PubMed] [Google Scholar]
  29. Naito A., Nagao T., Norisada K., Mizuno T., Tuzi S., Saitô H. Conformation and dynamics of melittin bound to magnetically oriented lipid bilayers by solid-state (31)P and (13)C NMR spectroscopy. Biophys J. 2000 May;78(5):2405–2417. doi: 10.1016/S0006-3495(00)76784-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Okada A., Wakamatsu K., Miyazawa T., Higashijima T. Vesicle-bound conformation of melittin: transferred nuclear Overhauser enhancement analysis in the presence of perdeuterated phosphatidylcholine vesicles. Biochemistry. 1994 Aug 16;33(32):9438–9446. doi: 10.1021/bi00198a009. [DOI] [PubMed] [Google Scholar]
  31. Rex S., Schwarz G. Quantitative studies on the melittin-induced leakage mechanism of lipid vesicles. Biochemistry. 1998 Feb 24;37(8):2336–2345. doi: 10.1021/bi971009p. [DOI] [PubMed] [Google Scholar]
  32. Schwarz G., Beschiaschvili G. Thermodynamic and kinetic studies on the association of melittin with a phospholipid bilayer. Biochim Biophys Acta. 1989 Feb 13;979(1):82–90. doi: 10.1016/0005-2736(89)90526-9. [DOI] [PubMed] [Google Scholar]
  33. Schwarz G., Zong R. T., Popescu T. Kinetics of melittin induced pore formation in the membrane of lipid vesicles. Biochim Biophys Acta. 1992 Sep 21;1110(1):97–104. doi: 10.1016/0005-2736(92)90299-2. [DOI] [PubMed] [Google Scholar]
  34. Selsted M. E., Novotny M. J., Morris W. L., Tang Y. Q., Smith W., Cullor J. S. Indolicidin, a novel bactericidal tridecapeptide amide from neutrophils. J Biol Chem. 1992 Mar 5;267(7):4292–4295. [PubMed] [Google Scholar]
  35. Sessa G., Freer J. H., Colacicco G., Weissmann G. Interaction of alytic polypeptide, melittin, with lipid membrane systems. J Biol Chem. 1969 Jul 10;244(13):3575–3582. [PubMed] [Google Scholar]
  36. Skehel J. J., Wiley D. C. Coiled coils in both intracellular vesicle and viral membrane fusion. Cell. 1998 Dec 23;95(7):871–874. doi: 10.1016/s0092-8674(00)81710-9. [DOI] [PubMed] [Google Scholar]
  37. Smith R., Separovic F., Milne T. J., Whittaker A., Bennett F. M., Cornell B. A., Makriyannis A. Structure and orientation of the pore-forming peptide, melittin, in lipid bilayers. J Mol Biol. 1994 Aug 19;241(3):456–466. doi: 10.1006/jmbi.1994.1520. [DOI] [PubMed] [Google Scholar]
  38. Strom R., Podo F., Crifo C., Berthet C., Zulauf M., Zaccai G. Structural aspects of the interaction of bee venom peptide melittin with phospholipids. Biopolymers. 1983 Jan;22(1):391–396. doi: 10.1002/bip.360220151. [DOI] [PubMed] [Google Scholar]
  39. Takei J., Remenyi A., Dempsey C. E. Generalised bilayer perturbation from peptide helix dimerisation at membrane surfaces: vesicle lysis induced by disulphide-dimerised melittin analogues. FEBS Lett. 1999 Jan 8;442(1):11–14. doi: 10.1016/s0014-5793(98)01617-2. [DOI] [PubMed] [Google Scholar]
  40. Terwilliger T. C., Eisenberg D. The structure of melittin. I. Structure determination and partial refinement. J Biol Chem. 1982 Jun 10;257(11):6010–6015. doi: 10.2210/pdb1mlt/pdb. [DOI] [PubMed] [Google Scholar]
  41. Terwilliger T. C., Eisenberg D. The structure of melittin. II. Interpretation of the structure. J Biol Chem. 1982 Jun 10;257(11):6016–6022. [PubMed] [Google Scholar]
  42. Terwilliger T. C., Weissman L., Eisenberg D. The structure of melittin in the form I crystals and its implication for melittin's lytic and surface activities. Biophys J. 1982 Jan;37(1):353–361. doi: 10.1016/S0006-3495(82)84683-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Vogel H., Jähnig F. The structure of melittin in membranes. Biophys J. 1986 Oct;50(4):573–582. doi: 10.1016/S0006-3495(86)83497-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. White S. H., Wimley W. C., Ladokhin A. S., Hristova K. Protein folding in membranes: determining energetics of peptide-bilayer interactions. Methods Enzymol. 1998;295:62–87. doi: 10.1016/s0076-6879(98)95035-2. [DOI] [PubMed] [Google Scholar]
  45. Wiener M. C., White S. H. Fluid bilayer structure determination by the combined use of x-ray and neutron diffraction. I. Fluid bilayer models and the limits of resolution. Biophys J. 1991 Jan;59(1):162–173. doi: 10.1016/S0006-3495(91)82208-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Wiener M. C., White S. H. Fluid bilayer structure determination by the combined use of x-ray and neutron diffraction. II. "Composition-space" refinement method. Biophys J. 1991 Jan;59(1):174–185. doi: 10.1016/S0006-3495(91)82209-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wiener M. C., White S. H. Structure of a fluid dioleoylphosphatidylcholine bilayer determined by joint refinement of x-ray and neutron diffraction data. III. Complete structure. Biophys J. 1992 Feb;61(2):434–447. doi: 10.1016/S0006-3495(92)81849-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wiener M. C., White S. H. Transbilayer distribution of bromine in fluid bilayers containing a specifically brominated analogue of dioleoylphosphatidylcholine. Biochemistry. 1991 Jul 16;30(28):6997–7008. doi: 10.1021/bi00242a027. [DOI] [PubMed] [Google Scholar]
  49. Wimley W. C., White S. H. Experimentally determined hydrophobicity scale for proteins at membrane interfaces. Nat Struct Biol. 1996 Oct;3(10):842–848. doi: 10.1038/nsb1096-842. [DOI] [PubMed] [Google Scholar]
  50. Wu Y., Huang H. W., Olah G. A. Method of oriented circular dichroism. Biophys J. 1990 Apr;57(4):797–806. doi: 10.1016/S0006-3495(90)82599-6. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES