Abstract
One promising strategy to combat the proliferation of bacteria resistance toward current antibiotics is the development of peptide-based drug. Among these compounds is a group of small cyclic peptides rich in arginine (Arg) and tryptophan (Trp) residues with selective toxicity toward Gram-negative bacteria. The small size of these peptides with improved toxicity toward Gram-negative bacteria makes them an interesting candidate to understand the forces responsible for their selectivity and paves the way to develop new therapeutics with potent activity toward multi-resistant Gram-negative bacteria. To reach this goal, isothermal titration calorimetry (ITC) is a useful technique which may provide the complete set of thermodynamic parameters of the interaction of peptides with lipid bilayers mimicking the properties of bacterial membranes within a few hours. The purpose of this chapter is to describe the synthesis of this group of small synthetic antimicrobial peptides together with the application of ITC to study their interaction with lipid membranes.
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References
Hancock, R. E. W. and Sahl, H. -G. (2006) Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nat. Biotechnol. 24, 1551–1557.
Chan, D. I., Prenner, E. J., and Vogel, H. J. (2006) Tryptophan- and arginine-rich antimicrobial peptides: structures and mechanisms of action. Biochim. Biophys. Acta 1758, 1184–1202.
Blondelle, S. E. and Houghten, R. A. (1996) Novel antimicrobial compounds identified using synthetic combinatorial library technology. Trends Biotechnol. 14, 60–65.
Dathe, M., Nikolenko, H., Klose, J., and Bienert, M. (2004) Cyclization increases the antimicrobial activity and selectivity of arginine- and tryptophan-containing hexapeptides. Biochemistry 43, 9140–9150.
Wessolowski, A., Bienert, M., and Dathe, M. (2004) Antimicrobial activity of arginine- and tryptophan-rich hexapeptides: the effects of aromatic clusters, D-amino acid substitution and cyclization. J. Pept. Res. 64, 159–169.
Appelt, C., Wessolowski, A., Soderhall, J. A., Dathe, M., and Schmieder, P. (2005) Structure of the antimicrobial, cationic hexapeptide cyclo (RRWWRF) and its analogues in solution and bound to detergent micelles. Chem. Bio. Chem. 6, 1654–1662.
Seelig, J. (1997) Titration of lipid-peptide interactions. Biochim. Biophys. Acta 1331, 103–116.
Ehrlich, A., Heyne, H. -U., Winter, R., Beyermann, M., Haber, H., Carpino, L. A., and Bienert, M. (1996) Cyclization of all-L-pentapeptides by means of 1-hydroxy-7-azabenzotriazole-derived uronium and phosphonium reagents. J. Org. Chem. 61, 8831–8838.
Ehrlich, A., Rothemund, S., Brudel, M., Beyermann, M., Carpino, L. A., and Bienert, M. (1993) Synthesis of cyclic peptides via efficient new coupling reagents. Tetrahedron Lett. 34, 4781–4784.
Bienert, M., Henklein, P., Beyermann, M., and Carpino, L. A. (2002) Uronium/guanidinium salts. In Synthesis of Peptides and Peptidomimetics (4th ed). Goodman M., Felix A., Moroder L., Toniolo C. (Eds.), pp. 555–580, Houben–Weyl Methods of Organic Chemistry. Stuttgart: Georg Thieme, Vol. E22a, Chapter 3.8.
Knorr, R., Trzeciak, A., Bannwarth, W., and Gillessen, D. (1989) New coupling reagents in peptide chemistry. Tetrahedron Lett. 30, 1927–1930.
Albericio, F., Bofill, J. M., El-Faham, A., and Kates, S. A. (1998) Use of onium salt-based coupling reagents in peptide synthesis. J. Org. Chem. 63, 9678–9683.
Thomas, P. G. and Seelig, J. (1993) Binding of the calcium antagonist flunarizine to phosphatidylcholine bilayers: charge effects and thermodynamics. Biochem. J. 291, 397–402.
Keller, S., Heerklotz, H., and Blume, A. (2006) Monitoring lipid membrane translocatlon of sodium dodecyl sulfate by isothermal titration calorimetry. J. Am. Chem. Soc. 128, 1279–1286.
Keller, S., Heerklotz, H., Jahnke, N., and Blume, A. (2006) Thermodynamics of lipid membrane solubilization by sodium dodecyl sulfate. Biophys J. 90, 4509–4521.
Acknowledgments
Dr. Sandro Keller (FMP) is gratefully acknowledged with ITC. Dr. Margitta Dathe (FMP) and Professor Michael Bienert (FMP) are thanked for their helpful comments on the manuscript.
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Bagheri, M. (2010). Synthesis and Thermodynamic Characterization of Small Cyclic Antimicrobial Arginine and Tryptophan-Rich Peptides with Selectivity for Gram-Negative Bacteria. In: Giuliani, A., Rinaldi, A. (eds) Antimicrobial Peptides. Methods in Molecular Biology, vol 618. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-594-1_7
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DOI: https://doi.org/10.1007/978-1-60761-594-1_7
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