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Synergistic Killing Effect of Synthetic Peptide P20 and Cefotaxime on Methicillin-Resistant Nosocomial Isolates of Staphylococcus aureus  

Jung, Hyun-Jun (Department of Microbiology, College of Natural Sciences, Kyungpook National University)
Choi, Kyu-Sik (Department of Microbiology, College of Natural Sciences, Kyungpook National University)
Lee, Dong-Gun (Department of Microbiology, College of Natural Sciences, Kyungpook National University)
Publication Information
Journal of Microbiology and Biotechnology / v.15, no.5, 2005 , pp. 1039-1046 More about this Journal
Abstract
The salt resistance of antibacterial activity and synergistic effect with clinically used antibiotic agents are critical factors in developing effective peptide antibiotic drugs. For this reason, we investigated the resistance of antibacterial activity to antagonism induced by NaCl and $MgCl_2$ and the synergistic effect of P20 with cefotaxime. P20 is a 20-residue synthetic peptide derived from a cecropin A (CA)-melittin(ME) hybrid peptide. In this study, P20 was found to have potent antibacterial activity against clinically isolated methicillin-resistant Staphylococcus aureus (MRSA) strains without hemolytic activity against human erythrocytes. The combination study revealed that P20 in combination with cefotaxime showed synergistic antibacterial activity in an energy-dependent manner. We also confirmed the synergism between P20 and cefotaxime by fluorescence-activated flow cytometric analysis by staining bacterial cells with propidium iodide (PI) and bis-(1,3-dibutylbarbituric acid) trimethine oxonol (BOX). This study suggests that P20 may be useful as a therapeutic antibiotic peptide with synergistic effect in combination with conventional antibiotic agents.
Keywords
Hybrid peptide; antibacterial activity; synergistic effect; cefotaxime; salts resistance; methicillin-resistant Staphylococcus aureus (MRSA);
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1 Park, P. J., J. Y. Je, H. G. Byun, S. H. Moon, and S. K Kim. 2004. Antimicrobial activity of hetero-chitosans and their oligosaccharides with different molecular weights. J. Microbiol. Biotechnol. 14: 317-323
2 Hayashi, H. and Y. Suzuki. 1998. Regulation of intracellular pH during H+-coupled oligopeptide absorption in enterocytes from guinea-pig ileum. J. Phys. 511: 573-586   DOI   ScienceOn
3 Simmaco, M., G. Mignogna, S. Canofeni, R. Miele, M. L. Mangoni, and D. Barra. 1996. Temporins, antimicrobial peptides from the European red frog Rana temporaria. Eur. J. Biochem. 242: 788- 792   DOI   ScienceOn
4 Park, Y, D. G. Lee, S. H. Jang, E. R. Woo, H. G. Jeong, D. H. Choi, and K. S. Hahm. 2003. A Leu-Lys-rich antimicrobial peptide: Activity and mechanism. Biochim. Biophys. Acta 1645: 172-182   DOI   ScienceOn
5 Giacometti, A., O. Cirioni, F. Barchiesi, M. Fortuna, and G. Scalise. 1999. In vitro activity of cationic peptides alone and in combination with clinically used antimicrobial agents against Pseudomonas aeruginosa. J. Antimicrob. Chemother. 44: 641-645   DOI
6 Lee, D. G, Y. S. Chang, Y. Park, K. S. Hahm, and E. R. Woo. 2002. Antimicrobial effects of ocotillone isolated from stem bark of Ailanthus altisshima. J. Microbiol. Biotechnol. 12: 854-857
7 Cormican, M. G. and R. N. Jones. 1996. Emerging resistance to antimicrobial agents in Gram-positive bacteria. Enterococci, Staphylococci and non pneumococcal streptococci. Drugs 51: S6-S12   DOI   ScienceOn
8 Lee, D. G., Y. Park, P. I. Kim, H. G. Jeong, E. R. Woo, and K. S. Hahm. 2002. Influence on the plasma membrane of Candida albicans by HP (2-9)-magainin 2 (1-12) hybrid peptide. Biochem. Biophys. Res. Commun. 297: 885-889   DOI   PUBMED   ScienceOn
9 Jungblut, P. and B. Thiede. 1997. Protein identification from 2-DE gels by MALDI mass spectrometry. Mass Spectrom. Rev. 16: 145-162   DOI   ScienceOn
10 Shin, S. Y., J. H. Kang, M. K. Lee, S. Y. Kim, Y. Kim, and K. S. Hahm. 1998. Cecropin A-magainin 2 hybrid peptides having potent antimicrobial activity with low hemolytic effect. Biochem. Mol. Biol. Int. 44: 1119-1126
11 Zasloff, M. 1987. Magainins, a class of antimicrobial peptides from Xenopus skin: Isolation characterization of two active forms, and partial cDNA sequence of precursor. Proc. Natl. Acad. Sci. USA 84: 5449-5453
12 Hancock, R. E. W. 1997. Peptides antibiotics. Lancet 349: 418-422   DOI   ScienceOn
13 Shin, S. Y., J. H. Kang, D. G. Lee, and K. S. Hahm. 1998. Design of novel antimicrobial peptides based on structure-antibiotic activity relationships of cecropin A, magainin 2 and melittin. J. Biochem. Mol. Biol. Biophys. 4: 135-145
14 Ganz, T. and R. I. Lehrer. 1995. Defensins. Pharmac. Ther. 66: 191-205   DOI   ScienceOn
15 Kim, D. H., D. G. Lee, K. L. Kim, and Y. Lee. 2001. Internalization of tenecin 3 by a fungal cellular process is essential for its fungicidal effect on Candida albicans. Eur. J. Biochem. 268: 4449-4458   DOI   ScienceOn
16 Houssin, C., D. T. Nguyen, G. Leblon, and N. Bayan. 2002. S-Iayer protein transport across the cell wall of Corynebacterium glutamicum: In vivo kinetics and energy requirements. FEMS Microbiol. Lett. 217: 71-79   DOI   ScienceOn
17 Leitch, E. C. and C. D. Willcox. 1999. Elucidation of the antistaphylococcal action of lactoferrin and lysozyme. J. Med. Microbiol. 48: 867-871   DOI   PUBMED
18 Werkmeister, J. A., A. Kirkpatrick, J. A. McKenzie, and D. E. Rivett. 1993. The effect of sequence variations and structure on the cytolytic activity of melittin peptides. Biochim. Biophys. Acta 1157: 50-54   DOI   ScienceOn
19 Sahl, H. G., R. W. Jack, and G. Bierbaum. 1995. Biosynthsis and biological activities of lantibiotics with unique post-translational modifications. Eur. J. Biochem. 230: 827-853   DOI   ScienceOn
20 Hewitt, C. J., R. Franke, A. Max, B. Kossmann, and P. Otterabach. 2004. A study into the anti-microbial properties of an amino functionalised polymer using multi-parameter flow cytometry. Biotechnol. Lett. 26: 549-557   DOI   ScienceOn
21 Conlon, J. M., A. Sonnerend, M. Patel, V. Camasamudram, N. Nowotny, E. Zilahi, S. Iwamuro, P. F. Nielsen, and T. Pal. 2003. A melittin-related peptide from the skin of the Japanese frog, Rana Tagoi, with antimicrobial and cytolytic properties. Biochem. Biophys. Res. Commun. 306: 496-500   DOI   ScienceOn
22 Labischinski, H., K. Ehlert, and B. Berger-Bachi. 1998. The targeting of factors necessary for expression of methicillin resistance in staphylococci. J. Antimicrob. Chemother. 41: 581-584   DOI   ScienceOn
23 Andreu, D., J. Ubach, A. Boman, B. Wahlin, D. Wade, R. B. Merrifield, and H. G. Boman. 1991. Shortened ceropin A-melittin hybrids: Significant size reduction retains potent antibiotics activity. FEBS Lett. 296: 190-194   DOI   ScienceOn
24 Broekaert, W., F. Terras, B. P. A. Cammue, and R. Osborne. 1995. Plant defensins: Novel antimicrobial peptides as components of the host defense system. Plant Physiol. 108: 1353-1358   DOI   ScienceOn
25 Hancock, R. E. W. and D. S. Chapple. 1999. Peptide antibiotics. Antimicrob. Agents Chemother. 43: 1317-1323
26 Shin, S. Y, M. K. Lee, K. L. Kim, and K. S. Hahm. 1997. Structure-antitumor and hemolytic activity relationships of synthetic peptides derived from cecropin A-magainin 2 and cecropin A-melittin hybrid peptides. J. Pept. Res. 50: 279¬285
27 Wade, D., A. Silveira, L. Rollins-Smith, T. Bergman, J. Silberring, and H. Lankinen. 2001. Hematological and antifungal properties of temporin A and a cecropin A-temporin A hybrid. Acta Biochim. Pol. 48: 1185-1189
28 Zasloff, M. 1992. Antibiotic peptides as mediators of innate immunity. Curr. Opin. Immunol. 4: 3-7   DOI   PUBMED   ScienceOn
29 Shin, S. Y, J. H. Kang, and K. S. Hahm. 1999. Structure-antibacterial, antitumor and hemolytic activity relationships of cecropin A-magainin 2 and cecropin A-melittin hybrid peptides. J. Pept. Res. 53: 82-90   DOI   ScienceOn
30 Park, P. J., H. K. Lee, and S. K. Kim. 2004. Preparation of hetero-chitooligosaccharides and their antimicrobial activity on Vibrio parahaemolyticus. J. Microbiol. Biotechnol. 14: 41-47
31 Mitsuhara, I. 2001. In vitro growth inhibition of human intestinal bacteria by sacrotoxin lA, an insect bactericidal peptide. Biotechnol. Lett. 23: 569-573   DOI   ScienceOn
32 Lee, D. G., Y. Park, K. S. Hahm, H. H. Lee, Y. H. Moon, and E. R. Woo. 2004. Structure-antiviral activity relationships of cecropin A-magainin 2 hybrid peptide and its analogues. J. Pept. Sci. 10: 298-303   DOI   ScienceOn
33 Dutta, B. P., S. C. Debnath, T. K. MandaI, and A. K. Chakraborty. 2004. Modification of pharmacokinetics of cefotaxime in uranyl nitrate-induced renal damage in black Bengal goats. J. Vet. Sci. 5: 1-3
34 Merrifield, R. B. 1986. Solid phase synthesis. Science 232: 341-347   DOI   PUBMED
35 Jack, R. W., J. R. Tagg, and B. Ray. 1995. Bacteriocins of Gram-positive bacteria. Microbiol. Rev. 59: 171-200
36 Kim, H. J., H. Peter Bennetto, and M. A. Halablab. 2004. Application of flow cytometry to monitoring of liposomal restructuring induced by Listeria monocytogenes. J. Microbiol. Biotechnol. 14: 1099-1102
37 Bals, R., X. Wang, M. Zasloff, and M. J. W. Wilson. 1998. The peptide antibiotic LL-37/hCAP-18 is expressed in epithelia of the human lung where it has broad antimicrobial activity at the airway surface. Proc. Natl. Acad. Sci. USA 95: 9541-9546