International Journal of Oral Biology

The Korean Academy of Oral Biology (대한구강생물학회)
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Identification of Antimicrobial Peptide Hexamers against Oral Pathogens through Rapid Screening of a Synthetic Combinatorial Peptide Library

  • Song, Je-Seon (Department of Pediatric Dentistry, Oral Science Research Center, College of Dentistry, Yonsei University) ;
  • Cho, Kyung Joo (Department of Oral Biochemistry and Molecular Biology, Kyung Hee University) ;
  • Kim, Joungmok (Department of Oral Biochemistry and Molecular Biology, Kyung Hee University) ;
  • Kim, Jeong Hee (Department of Oral Biochemistry and Molecular Biology, Kyung Hee University)
  • Received : 2014.07.02
  • Accepted : 2014.08.25
  • Published : 2014.12.31
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Abstract

A positional scanning synthetic peptide combinatorial library (PS-SCL) was screened in order to identify antimicrobial peptides against the cariogenic oral bacteria, Streptococcus mutans. Activity against Streptococcus gordonii and Aggregatibacter actinomycetemcomitans was also examined. The library was comprised of six sub-libraries with the format $O_{(1-6)}XXXXX-NH_2$, where O represents one of 19 amino acids (excluding cysteine) and X represents equimolar mixture of these. Each sub-library was tested for antimicrobial activity against S. mutans and evaluated for antimicrobial activity against S. gordonii and A. actinomycetemcomitans. The effect of peptides was observed using transmission electron microscopy (TEM). Two semi-mixture peptides, RXXXXN-$NH_2$ (pep-1) and WXXXXN-$NH_2$ (pep-2), and one positioned peptide, RRRWRN-$NH_2$ (pep-3), were identified. Pep-1 and pep-2 showed significant antimicrobial activity against Gram positive bacteria (S. mutans and S. gordonii), but not against Gram negative bacteria (A. actinomycetemcomitans). However, pep-3 showed very low antimicrobial activity against all three bacteria. Pep-3 did not form an amphiphilic ${\alpha}$-helix, which is a required structure for most antimicrobial peptides. Pep-1 and pep-2 were able to disrupt the membrane of S. mutans. Small libraries of biochemically-constrained peptides can be used to generate antimicrobial peptides against S. mutans and other oral microbes. Peptides derived from such libraries may be candidate antimicrobial agents for the treatment of oral microorganisms.

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References

  1. Hamada S, Slade HD. Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev. 1980;44:331-384.
  2. Loesche WJ. Role of Streptococcus mutans in human dental decay. Microbiol Rev. 1986;50:353-380.
  3. Loesche WJ. The identification of bacteria associated with periodontal disease and dental caries by enzymatic methods. Oral Microbiol Immunol. 1986;1:65-72. https://doi.org/10.1111/j.1399-302X.1986.tb00322.x
  4. Griffen AL, Becker MR, Lyons SR, Moeschberger ML, Leys EJ. Prevalence of Porphyromonas gingivalis and periodontal health status. J Clin Microbiol. 1998;36:3239-3242.
  5. Slots J, Ting M. Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in human periodontal disease: occurrence and treatment. Periodontol 2000 1999;20:82-121. https://doi.org/10.1111/j.1600-0757.1999.tb00159.x
  6. Rolla G, Melsen B. On the mechanism of the plaque inhibition by chlorhexidine. J Dent Res. 1975;54:B57-62. https://doi.org/10.1177/00220345750540022601
  7. De Paola PF, Jordan HV, Berg J. Temporary suppression of Streptococcus mutans in humans through topical application of vancomycin. J Dent Res. 1974;53:108-114. https://doi.org/10.1177/00220345740530010201
  8. Englander HR, Keyes PH. Control of Streptococcus mutans, plaque, and dental caries in hamsters with topically applied vancomycin. Arch Oral Biol. 1971;16:469-472. https://doi.org/10.1016/0003-9969(71)90171-3
  9. Brown AT, Largent BA, Ferretti GA, Lillich TT. Chemical control of plaque-dependent oral diseases: the use of chlorhexidine. Compendium 1986;7:719-720, 722-724.
  10. Flotra L, Gjermo P, Rolla G, Waerhaug J. Side effects of chlorhexidine mouth washes. Scand J Dent Res. 1971;79:119-125.
  11. Davies J. Inactivation of antibiotics and the dissemination of resistance genes. Science 1994;264:375-382. https://doi.org/10.1126/science.8153624
  12. Boman HG. Peptide antibiotics and their role in innate immunity. Annu Rev Immunol. 1995;13:61-92. https://doi.org/10.1146/annurev.iy.13.040195.000425
  13. Hancock RE, Sahl HG. Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nat Biotechnol. 2006;24:1551-1557. https://doi.org/10.1038/nbt1267
  14. Jenssen H, Hamill P, Hancock RE. Peptide antimicrobial agents. Clin Microbiol Rev. 2006 ;19:491-511. https://doi.org/10.1128/CMR.00056-05
  15. Zasloff M. Antimicrobial peptides of multicellular organisms. Nature 2002;415:389-395. https://doi.org/10.1038/415389a
  16. Brogden KA. Antimicrobial peptides: pore formers or metabolic inhibitors in bacteria? Nat Rev Microbiol. 2005;3:238-250. https://doi.org/10.1038/nrmicro1098
  17. Epand RM, Vogel HJ. Diversity of antimicrobial peptides and their mechanisms of action. Biochim Biophys Acta. 1999;1462:11-28. https://doi.org/10.1016/S0005-2736(99)00198-4
  18. Bals R. Epithelial antimicrobial peptides in host defense against infection. Respir Res. 2000;1:141-150. https://doi.org/10.1186/rr25
  19. Hancock RE, Diamond G. The role of cationic antimicrobial peptides in innate host defences. Trends Microbiol. 2000;8:402-410. https://doi.org/10.1016/S0966-842X(00)01823-0
  20. Mookherjee N, Hancock RE. Cationic host defence peptides: innate immune regulatory peptides as a novel approach for treating infections. Cell Mol Life Sci. 2007;64:922-933. https://doi.org/10.1007/s00018-007-6475-6
  21. Koczulla AR, Bals R. Antimicrobial peptides: current status and therapeutic potential. Drugs 2003;63:389-406. https://doi.org/10.2165/00003495-200363040-00005
  22. Houghten RA, Pinilla C, Blondelle SE, Appel JR, Dooley CT, Cuervo JH. Generation and use of synthetic peptide combinatorial libraries for basic research and drug discovery. Nature 1991;354:84-86. https://doi.org/10.1038/354084a0
  23. Lee KH. Development of short antimicrobial peptides derived from host defense peptides or by combinatorial libraries. Curr Pharm Des. 2002;8:795-813. https://doi.org/10.2174/1381612023395411
  24. He J, Eckert R, Pharm T, Simanian MD, Hu C, Yarbrough DK, Qi F, Anderson MH, Shi W. Novel synthetic antimicrobial peptides against Streptococcus mutans. Antimicrob Agents Chemother. 2007;51:1351-1358. https://doi.org/10.1128/AAC.01270-06
  25. Kim SS, Kim S, Kim E, Hyun B, Kim KK, Lee BJ. Synergistic inhibitory effect of cationic peptides and antimicrobial agents on the growth of oral streptococci. Caries Res. 2003;37:425-430. https://doi.org/10.1159/000073394
  26. Ryge TS, Hansen PR. Potent antibacterial lysine-peptoid hybrids identified from a positional scanning combinatorial library. Bioorg Med Chem. 2006;14:4444-4451. https://doi.org/10.1016/j.bmc.2006.02.034
  27. Denholt CL, Hansen PR, Pedersen N, Poulsen HS, Gillings N, Kjaer A. Identification of novel peptide ligands for the cancer-specific receptor mutation EFGRvIII using a mixture-based synthetic combinatorial library. Biopolymers 2009;91:201-206. https://doi.org/10.1002/bip.21117
  28. Schmid B, Warnecke A, Fichtner I, Jung M, Kratz F. Development of albumin-binding camptothecin prodrugs using a Peptide positional scanning library. Bioconjug Chem. 2007;18:1786-1799. https://doi.org/10.1021/bc0700842
  29. Choi J, Moon E. Identification of novel bioactive hexapeptides against phytopathogenic bacteria through rapid screening of a synthetic combinatorial library. J Microbiol Biotechnol. 2009;19:792-802. https://doi.org/10.4014/jmb.0809.497
  30. Armishaw CJ, Singh N, Medina-Franco JL, Clark RJ, Scott KC, Houghten RA, Jensen AA. A synthetic combinatorial strategy for developing a-conotoxin analogs as potent a7 nicotinic acetylcholine receptor antagonists. J Biol Chem. 2010;285:1809-1821. https://doi.org/10.1074/jbc.M109.071183
  31. Armishaw CJ, Banerjee J, Ganno ML, Reilley KJ, Eans SO, Mizrachi E, Gyanda R, Hoot MR, Houghten RA, McLaughlin JP. Discovery of novel antinociceptive ${\alpha}$-conotoxin analogues from the direct in vivo screening of a synthetic mixture-based combinatorial library. ACS Comb Sci. 2013;15:153-161. https://doi.org/10.1021/co300152x
  32. Lam KS, Salmon SE, Hersh EM, Hruby VJ, Kazmierski WM, Knapp RJ. A new type of synthetic peptide library for identifying ligand-binding activity. Nature 1991;354:82-84. https://doi.org/10.1038/354082a0
  33. Hancock RE, Rozek A. Role of membranes in the activities of antimicrobial cationic peptides. FEMS Microbiol Lett. 2002;206:143-149. https://doi.org/10.1111/j.1574-6968.2002.tb11000.x
  34. Matsuzaki K. Why and how are peptide-lipid interactions utilized for self-defense? Magainins and tachyplesins as archetypes. Biochim Biophys Acta. 1999;1462:1-10. https://doi.org/10.1016/S0005-2736(99)00197-2
  35. Wang K, Yan J, Dang W, Liu X, Chen R, Zhang J, Zhang B, Zhang W, Kai M, Yan W, Yang Z, Xie J, Wang R. Membrane active antimicrobial activity and molecular dynamics study of a novel cationic antimicrobial peptide polybia-MPI, from the venom of Polybia paulista. Peptides 2013;39:80-88. https://doi.org/10.1016/j.peptides.2012.11.002
  36. Wiradharma N, Khoe U, Hauser CAE, Seow SV, Zhang S, Yang Y. Synthetic cationic amphiphilic ${\alpha}$-helical peptides as antimicrobial agents. Biomaterials 2011;32:2204-2212. https://doi.org/10.1016/j.biomaterials.2010.11.054
  37. Douglas CW, Heath J, Hampton KK, Preston FE. Identity of viridans streptococci isolated from cases of infective endocarditis. J Med Microbiol. 1993;39:179-182. https://doi.org/10.1099/00222615-39-3-179
  38. Durak DT, Phil MBD. Prevention of infective endocarditis. N Engl J Med. 1995;332:38-44. https://doi.org/10.1056/NEJM199501053320107
  39. Faveri M, figueiredo LC, Duarte PM, Mestnik MJ, Mayer MP, Feres M. Microbiological profiles of untreated subjects with localized aggressive periodontitis. J Clin Periodontol. 2009;36:739-749. https://doi.org/10.1111/j.1600-051X.2009.01449.x