Summary
Widespread resistance to antibiotics in current clinical use is increasing at an alarming rate. Novel approaches in antimicrobial therapy will be required in the near future to maintain control of infectious diseases. An enormous array of small cationic peptides exists in nature as part of the innate defense systems of organisms ranging from bacteria to humans. For most naturally occurring linear peptides, such as magainins and cecropins, a common feature is their capacity to form an amphipathic α-helix (with polar and nonpolar groups on opposite faces of the helix), a structural feature believed to be important in their antimicrobial function as membrane-lytic agents. A massive effort over the past two decades has resulted in a better understanding of the molecular mechanism of antimicrobial peptides and the production of more potent analogues. To date, however, few of these peptides have been shown to have clinical efficacy, especially for systemic use, in large part due to insufficient selectivity between target and host cells. Recently, we developed a new strategy in the design of antimicrobial peptides. These linear cationic peptides, which form amphipathic β-sheets rather than α-helices, demonstrated superior selectivity in binding to the lipids contained in bacterial vs. mammalian plasma membranes. Here we describe methods to evaluate the structure and function of cationic antimicrobial peptides.
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References
Blazyk, J., Wiegand, R., Klein, J., Hammer, J., Epand, R. M., Epand, R. F, Maloy, W. L., and Kari, U. P. (2001) A novel linear amphipathic beta-sheet cationic antimicrobial peptide with enhanced selectivity for bacterial lipids. J. Biol. Chem. 276, 27899–27906.
Jin, Y., Mozsolits, H., Hammer, J., Zmuda, E., Zhu, F., Zhang, Y., Aguilar, M. I., and Blazyk, J. (2003) Influence of tryptophan on lipid binding of linear amphipathic cationic antimicrobial peptides. Biochemistry 42, 9395–9405.
Jin, Y., Hammer, J., Pate, M., Zhang, Y., Zhu, F., Zmuda, E., and Blazyk, J. (2005) Antimicrobial activities and structures of two linear cationic peptide families with various amphipathic beta-sheet and alpha-helical potentials. Antimicrob. Agents Chemother. 49, 4957–4964.
Skerlavaj, B., Romeo, D., and Gennaro, R. (1990) Rapid membrane permeabilization and inhibition of vital functions of Gram-negative bacteria by bactenecins. Infect. Immun. 58, 3724–3730.
Lakowicz, J. R. (1999) Protein fluorescence. In Principles of Fluorescence Spectroscopy, 2nd ed. Kluwer Academic/Plenum Publishers, New York, pp. 445–486.
Breukink, E., Van Kraaij, C., Van Dalen, A., Demel, R. A., Siezen, R. J., De Kruijff, B., and Kuipers, O. P. (1998) The orientation of nisin in membranes. Biochemistry 37, 8153–8162.
Schibli, D. J., Hwang, P. M., and Vogel, H. J. (1999) Structure of the antimicrobial peptide tritrpticin bound to micelles: A distinct membrane-bound peptide fold. Biochemistry 38, 16749–16755.
Ladokhin, A. S. (1999) Analysis of protein and peptide penetration into membranes by depth-dependent fluorescence quenching: Theoretical considerations. Biophys. J. 76, 946–955.
Epand, R. F., Epand, R. M., Monaco, V., Stoia, S., Formaggio, F., Crisma, M., and Toniolo, C. (1999) The antimicrobial peptide trichogin and its interaction with phospholipid membranes. Eur. J. Biochem. 266, 1021–1028.
Hong, J., Oren, Z., and Shai, Y. (1999) Structure and organization of hemolytic and nonhemolytic diastereomers of antimicrobial peptides in membranes. Biochemistry 38, 16963–16973.
Campbell, I. D., and Dwek, R. A. (1984) Optical activity. In Biological Spectroscopy. The Benjamin/Cummings Publishing Company, Inc., Menlo Park, CA, pp. 255–277.
Chen, P. S., Toribara, T. Y., and Warner, H. (1956) Microdetermination of phosphorus. Anal. Chem. 28, 1756–1758.
Ladokhin, A. S., Jayasinghe, S., and White, S. H. (2000) How to measure and analyze tryptophan fluorescence in membranes properly, and why bother? Anal. Biochem. 285, 235–245.
Acknowledgments
The authors would like to thank Dr. Renato Gennaro for providing the E. coli ML-35 bacterial strain and Dr. Alexey Ladokhin for helpful discussions concerning the fluorescence experiments. This work was supported by NIH Grants AI-047165 and C06-RR-14575 by the Ohio University College of Osteopathic Medicine.
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Pate, M., Blazyk, J. (2008). Methods for Assessing the Structure and Function of Cationic Antimicrobial Peptides. In: Champney, W.S. (eds) New Antibiotic Targets. Methods In Molecular Medicine™, vol 142. Humana Press. https://doi.org/10.1007/978-1-59745-246-5_13
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DOI: https://doi.org/10.1007/978-1-59745-246-5_13
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