European Biophysics Journal

, Volume 41, Issue 7, pp 629–636 | Cite as

NMR studies on the monomer–tetramer transition of melittin in an aqueous solution at high and low temperatures

Original Paper

Abstract

Melittin, a peptide of 26 amino acid residues, has been used as a model peptide for protein folding and unfolding, and extensive research has been done into its structure and conformational stability. Circular dichroism (CD) studies have demonstrated that melittin in an aqueous solution undergoes a transition from a helical tetramer to a random coil monomer not only by heating but also by cooling from room temperature (i.e., heat- and cold-denaturation, respectively). The heat-denaturation has been also examined by nuclear magnetic resonance (NMR) experiments, however, no NMR data have been presented on the cold-denaturation. In this paper, using proton (1H) NMR spectroscopy, we show that melittin undergoes conformational transitions from the monomer to the tetramer to the monomer by elevating temperature from 2 to 70 °C. Only melittin including a trans proline peptide bond participates in the transitions, whereas melittin including a cis proline one does not. The tetramer has maximum conformation stability at around 20 °C, and cooperativity of the heat-denaturation is extremely low.

Keywords

Melittin Self-association Monomer–tetramer transition NMR spectroscopy cis and trans proline peptide bonds Cold- and heat-denaturation 

References

  1. Bazzo R, Tappin MJ, Pastore A, Harvey TS, Carver JA, Campbell ID (1988) The structure of melittin: a 1H-NMR study in methanol. Eur J Biochem 173:139–146PubMedCrossRefGoogle Scholar
  2. Bello J, Bello HR, Granados E (1982) Conformation and aggregation of melittin: dependence on pH and concentration. Biochemistry 21:461–465PubMedCrossRefGoogle Scholar
  3. Brown LR, Lauterwein J, Wüthrich K (1980) High–resolution 1H-NMR studies of self-aggregation of melittin in aqueous solution. Biochim Biophys Acta 622:231–244PubMedCrossRefGoogle Scholar
  4. Chandler D (2005) Interfaces and the driving force of hydrophobic assembly. Nature 437:640–647PubMedCrossRefGoogle Scholar
  5. Dempsey CE (1990) The action of melittin on membranes. Biochim Biophys Acta 1031:143–230PubMedCrossRefGoogle Scholar
  6. Dias CL, Ala-Nissila T, Karttunen M, Vattulainen I, Grant M (2008) Microscopic mechanism for cold denaturation. Phys Rev Lett 21:118101CrossRefGoogle Scholar
  7. Faucon JF, Dufourco J, Lussan C (1979) The self-association of melittin and its binding to lipids. FEBS Lett 102:187–190PubMedCrossRefGoogle Scholar
  8. Goto Y, Hagihara Y (1992) Mechanism of the conformational transition of melittin. Biochemistry 31:732–738PubMedCrossRefGoogle Scholar
  9. Hagihara Y, Kataoka M, Aimoto S, Goto Y (1992) Charge repulsion in the conformational stability of melittin. Biochemistry 31:11908–11914PubMedCrossRefGoogle Scholar
  10. Hagihara Y, OObatake M, Goto Y (1994) Thermal unfolding of tetramer melittin: comparison with the molten golobule state of cytochrome c. Protein Sci 3:1418–1429PubMedCrossRefGoogle Scholar
  11. Inagaki F, Shimada I, Kawaguchi K, Hirano M, Terasawa I, Ikura T, Go N (1989) Structure of melittin bound to perdeuterated dodecylphosphocholine micelles as studied by two-dimensional NMR and distant geometry calculations. Biochemistry 28:5985–5991CrossRefGoogle Scholar
  12. Iwadate M, Asakura T, Williamson MP (1998) The structure of the melittin at different temperatures: an NOE-based calculation with chemical shift refinement. Eur J Biochem 257:479–487PubMedCrossRefGoogle Scholar
  13. Kinoshita M (2009) Importance of translational entropy of water in biological self-assembly processes like protein folding. Int J Mol Sci 10:1064–1080PubMedCrossRefGoogle Scholar
  14. Lauterwein J, Brown LR, Wüthrich K (1980) High-resolution 1H-NMR studies of monomeric melittin in aqueous solution. Biochim Biophys Acta 622:219–230PubMedCrossRefGoogle Scholar
  15. Marques MI, Borreguero JM, Stanley HE, Dokholyan NV (2003) Possible mechanism for cold denaturation of proteins at high temperature. Phys Rev Lett 91:138103PubMedCrossRefGoogle Scholar
  16. Miura Y (2011) Helix conformation of a small peptide melittin in a methanol-water mixed solvent studied by NMR. Protein Pept Lett 18:318–326PubMedCrossRefGoogle Scholar
  17. Murphy KP, Privalov PL, Gill SJ (1990) Common features of protein unfolding and dissolution of hydrophobic compounds. Science 247:559–561PubMedCrossRefGoogle Scholar
  18. Othon CM, Kwon O, Lin MM, Zewail AH (2009) Solvation in protein (un)folding of melittin tetramer-monomer transition. PNAS 106:12593–12598PubMedCrossRefGoogle Scholar
  19. Privalov PL (1990) Cold denaturation of proteins. Crit Rev Biochem Mol Biol 25:281–306PubMedCrossRefGoogle Scholar
  20. Privalov PL (1992) Physical basis of the stability of the folded conformations of proteins. In: Creighton TE (ed) Protein folding. Freeman WE and Company, New York, pp 83–126Google Scholar
  21. Privalov PL, Gill SJ (1988) Stability of protein structure and hydrophobic interaction. Adv Protein Chem 39:191–234PubMedCrossRefGoogle Scholar
  22. Qiu W, Zhang L, Kao Y, Lu W, Li T, Kim J, Sollenberger GM, Wang L, Zhong D (2005) Ultra hydration dynamics in melittin folding and aggregation: helix formation and tetramer self-assembly. J Phys Chem B 109:16901–16910PubMedCrossRefGoogle Scholar
  23. Quay SC, Condie CC (1983) Conformational studies of aqueous melittin: thermodynamic parameters of the monomer–tetramer self-association reaction. Biochemistry 22:695–700PubMedCrossRefGoogle Scholar
  24. Ramalingam K, Bello J, Aimoto S (1991) Conformation changes in melittin upon complexation with an anionic melittin analog. FEBS Lett 295:200–202PubMedCrossRefGoogle Scholar
  25. Ramalingam K, Aimoto S, Bello J (1992) Conformational studies of anionic melittin analogues: effect of peptide concentration, pH, ionic strength, and temperature—models for protein folding and halophilic proteins. Biopolymers 32:981–992PubMedCrossRefGoogle Scholar
  26. Schubert D, Pappert G, Boss K (1985) Does dimeric melittin occur in aqueous solutions? Biophys J 48:327–329PubMedCrossRefGoogle Scholar
  27. Soda K (1993) Structural and thermodynamic aspects of the hydrophobic effect. Adv Biophys 29:1–54PubMedCrossRefGoogle Scholar
  28. ten Wolde PR, Chandler D (2002) Drying-induced hydrophobic polymer collapse. Proc Natl Acad Sci USA 99:6539–6543PubMedCrossRefGoogle Scholar
  29. Wilcox W, Eisenberg D (1992) Thermodynamics of melittin tetramerization determined by circular dichroism and implications for protein folding. Protein Sci 1:641–653PubMedCrossRefGoogle Scholar
  30. Yoshidome T, Kinoshita M (2009) Hydrophobicity at low temperatures and cold denaturation of a protein. Phys Rev E 79:030905(R)Google Scholar

Copyright information

© European Biophysical Societies' Association 2012

Authors and Affiliations

  1. 1.Center for Advanced Instrumental AnalysisKyushu UniversityKasugaJapan

Personalised recommendations