Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Effects of C-Terminal Residues of 12-Mer Peptides on Antibacterial Efficacy and Mechanism

  • Son, Kkabi (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Kim, Jieun (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Jang, Mihee (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Chauhan, Anil Kumar (Department of Bioscience and Biotechnology, Konkuk University) ;
  • Kim, Yangmee (Department of Bioscience and Biotechnology, Konkuk University)
  • Received : 2019.07.26
  • Accepted : 2019.09.11
  • Published : 2019.11.28
Download PDF
⟨ Previous Next ⟩

Abstract

The development of new antimicrobial agents is essential for the effective treatment of diseases such as sepsis. We previously developed a new short peptide, Pap12-6, using the 12 N-terminal residues of papiliocin, which showed potent and effective antimicrobial activity against multidrug-resistant Gram-negative bacteria. Here, we investigated the antimicrobial mechanism of Pap12-6 and a newly designed peptide, Pap12-7, in which the 12th Trp residue of Pap12-6 was replaced with Val to develop a potent peptide with high bacterial selectivity and a different antibacterial mechanism. Both peptides showed high antimicrobial activity against Gram-negative bacteria, including multidrug-resistant Gram-negative bacteria. In addition, the two peptides showed similar anti-inflammatory activity against lipopolysaccharide-stimulated RAW 264.7 cells, but Pap12-7 showed very low toxicities against sheep red blood cells and mammalian cells compared to that showed by Pap12-6. A calcein dye leakage assay, membrane depolarization, and confocal microscopy observations revealed that the two peptides with one single amino acid change have different mechanisms of antibacterial action: Pap12-6 directly targets the bacterial cell membrane, whereas Pap12-7 appears to penetrate the bacterial cell membrane and exert its activities in the cell. The therapeutic efficacy of Pap12-7 was further examined in a mouse model of sepsis, which increased the survival rate of septic mice. For the first time, we showed that both peptides showed anti-septic activity by reducing the infiltration of neutrophils and the production of inflammatory factors. Overall, these results indicate Pap12-7 as a novel non-toxic peptide with potent antibacterial and anti-septic activities via penetrating the cell membrane.

Keywords

References

  1. Carlet J, Collignon P, Goldmann D, Goossens H, Gyssens IC, Harbarth S, et al. 2011. Society's failure to protect a precious resource: antibiotics. Lancet 378: 369-371. https://doi.org/10.1016/S0140-6736(11)60401-7
  2. Bragginton EC, Piddock LJ. 2014. UK and European Union public and charitable funding from 2008 to 2013 for bacteriology and antibiotic research in the UK: an observational study. Lancet Infect. Dis. 14: 857-868. https://doi.org/10.1016/s1473-3099(14)70825-4
  3. Kostyanev T, Bonten MJ, O'Brien S, Steel H, Ross S, Francois B, et al. 2016. The innovative medicines initiative's new drugs for bad bugs programme: European public-private partnerships for the development of new strategies to tackle antibiotic resistance. J. Antimicrob. Chemother. 71: 290-295. https://doi.org/10.1093/jac/dkv339
  4. Karam G, Chastre J, Wilcox MH, Vincent JL. 2016. Antibiotic strategies in the era of multidrug resistance. Crit. Care. 20(1): 136. https://doi.org/10.1186/s13054-016-1320-7
  5. Medina E, Pieper DH. 2016. Tackling threats and future problems of multidrug-resistant bacteria. Curr. Top. Microbiol. Immunol. 398: 3-33.
  6. Godreuil S, Leban N, Padilla A, Hamel R, Luplertlop N, Chauffour A, et al. 2014. Aedesin: structure and antimicrobial activity against multidrug resistant bacterial strains. PLoS One 9: e105441. https://doi.org/10.1371/journal.pone.0105441
  7. Haney EF, Mansour SC, Hancock RE. 2017. Antimicrobial peptides: An introduction. Methods Mol. Biol. 1548: 3-22. https://doi.org/10.1007/978-1-4939-6737-7_1
  8. da Costa JP, Cova M, Ferreira R, Vitorino R. 2015. Antimicrobial peptides: an alternative for innovative medicines? Appl. Microbiol. Biotechnol. 99: 2023-2040. https://doi.org/10.1007/s00253-015-6375-x
  9. Kokel A, Torok M. 2018. Recent advances in the development of antimicrobial peptides (AMPs): Attempts for sustainable medicine? Curr. Med. Chem. 25: 2503-2519. https://doi.org/10.2174/0929867325666180117142142
  10. Galdiero S, Falanga A, Cantisani M, Vitiello M, Morelli G, Galdiero M. 2013. Peptide-lipid interactions: experiments and applications. Int. J. Mol. Sci. 14: 18758-18789. https://doi.org/10.3390/ijms140918758
  11. Kristensen M, Birch D, Morck Nielsen H. 2016. Applications and challenges for use of cell-penetrating peptides as delivery vectors for peptide and protein cargos. Int. J. Mol. Sci. 17(2). pii: E185. doi: 10.3390/ijms17020185.
  12. Guidotti G, Brambilla L, Rossi D. 2017. Cell-Penetrating Peptides: from basic research to clinics. Trends Pharmacol. Sci. 38: 406-424. https://doi.org/10.1016/j.tips.2017.01.003
  13. Uematsu N, Matsuzaki K. 2000. Polar angle as a determinant of amphipathic alpha-helix-lipid interactions: a model peptide study. Biophys. J. 79: 2075-2083. https://doi.org/10.1016/S0006-3495(00)76455-1
  14. Yeaman MR, Yount NY. 2003. Mechanisms of antimicrobial peptide action and resistance. Pharmacol. Rev. 55: 27-55. https://doi.org/10.1124/pr.55.1.2
  15. Lee TH, Hall KN, Aguilar MI. 2016. Antimicrobial peptide structure and mechanism of action: A focus on the role of membrane structure. Curr. Top. Med. Chem. 16: 25-39. https://doi.org/10.2174/1568026615666150703121700
  16. Kumar P, Kizhakkedathu JN, Straus SK. 2018. Antimicrobial peptides: Diversity, mechanism of action and strategies to improve the activity and biocompatibility in vivo. Biomolecules. 8(1). pii: E4. doi: 10.3390/biom8010004.
  17. Lee E, Jeong KW, Lee J, Shin A, Kim JK, Lee J, et al. 2013. Structure-activity relationships of cecropin-like peptides and their interactions with phospholipid membrane. BMB Rep. 46: 282-287. https://doi.org/10.5483/BMBRep.2013.46.5.252
  18. Wu Q, Patocka J, Kuca K. 2018. Insect Antimicrobial peptides, a Mini Review. Toxins (Basel). 10(11). pii: E461.
  19. Henriques ST, Melo MN, Castanho MA. 2006. Cell-penetrating peptides and antimicrobial peptides: how different are they? Biochem. J. 399: 1-7. https://doi.org/10.1042/BJ20061100
  20. Kim JK, Lee E, Shin S, Jeong KW, Lee JY, Bae SY, et al. 2011. Structure and function of papiliocin with antimicrobial and anti-inflammatory activities isolated from the swallowtail butterfly, Papilio xuthus. J. Biol. Chem. 286: 41296-41311. https://doi.org/10.1074/jbc.M111.269225
  21. Jeon D, Jacob B, Kwak C, Kim Y. 2017. Short antimicrobial peptides exhibiting antibacterial and anti-inflammatory activities derived from the N-terminal helix of papiliocin. B. Korean Chem. Soc. 38: 1260-1268. https://doi.org/10.1002/bkcs.11277
  22. Kim J, Jacob B, Jang M, Kwak C, Lee Y, Son K, et al. 2019. Development of a novel short 12-meric papiliocin-derived peptide that is effective against Gram-negative sepsis. Sci. Rep. 9: 3817. https://doi.org/10.1038/s41598-019-40577-8
  23. Ma QQ, Shan AS, Dong N, Cao YP, Lv YF, Wang L. 2011. The effects of Leu or Val residues on cell selectivity of alpha-helical peptides. Protein Pept. Lett. 18: 1112-1118. https://doi.org/10.2174/092986611797200968
  24. Bea Rde L, Petraglia AF, Johnson LE. 2015. Synthesis, antimicrobial activity and toxicity of analogs of the scorpion venom BmKn peptides. Toxicon. 101: 79-84. https://doi.org/10.1016/j.toxicon.2015.05.006
  25. Lee E, Kim JK, Jeon D, Jeong KW, Shin A, Kim Y. 2015. Functional roles of aromatic residues and helices of papiliocin in its antimicrobial and anti-inflammatory activities. Sci. Rep. 5: 12048. https://doi.org/10.1038/srep12048
  26. Lee E, Shin A, Kim Y. 2015. Anti-inflammatory activities of cecropin A and its mechanism of action. Arch. Insect. Biochem. Physiol. 88: 31-44. https://doi.org/10.1002/arch.21193
  27. Jeon D, Jeong MC, Jacob B, Bang JK, Kim EH, Cheong C, et al. 2017. Investigation of cationicity and structure of pseudin-2 analogues for enhanced bacterial selectivity and antiinflammatory activity. Sci. Rep. 7: 1455. 28 .
  28. Jnawali HN, Lee E, Jeong KW, Shin A, Heo YS, Kim Y. 2014. Anti-inflammatory activity of rhamnetin and a model of its binding to c-Jun NH2-terminal kinase 1 and p38 MAPK. J. Nat. Prod. 77: 258-263. https://doi.org/10.1021/np400803n
  29. Jnawali HN, Jeon D, Jeong MC, Lee E, Jin B, Ryoo S, et al. 2016. Antituberculosis Activity of a Naturally Occurring Flavonoid, Isorhamnetin. J. Nat. Prod. 79: 961-969. https://doi.org/10.1021/acs.jnatprod.5b01033
  30. Moussa HG, Martins AM, Husseini GA. 2015. Review on triggered liposomal drug delivery with a focus on ultrasound. Curr. Cancer. Drug Targets 15: 282-313. https://doi.org/10.2174/1568009615666150311100610
  31. Watson H. 2015. Biological membranes. Essays Biochem. 59: 43-69. https://doi.org/10.1042/bse0590043
  32. Li J, Koh JJ, Liu S, Lakshminarayanan R, Verma CS, Beuerman RW. 2017. Membrane active antimicrobial peptides: Translating mechanistic insights to design. Front. Neurosci. 11: 73.
  33. Epand RM, Epand RF. 2009. Lipid domains in bacterial membranes and the action of antimicrobial agents. Biochim. Biophys. Acta 1788: 289-294. https://doi.org/10.1016/j.bbamem.2008.08.023
  34. Dowhan W. 1997. Molecular basis for membrane phospholipid diversity: why are there so many lipids? Annu. Rev. Biochem. 66: 199-232. https://doi.org/10.1146/annurev.biochem.66.1.199
  35. Mingeot-Leclercq MP, Decout JL. 2016. Bacterial lipid membranes as promising targets to fight antimicrobial resistance, molecular foundations and illustration through the renewal of aminoglycoside antibiotics and emergence of amphiphilic aminoglycosides. Medchemcomm 7: 586-611. https://doi.org/10.1039/c5md00503e
  36. Sohlenkamp C, Geiger O. 2016. Bacterial membrane lipids: diversity in structures and pathways. FEMS Microbiol. Rev. 40: 133-159. https://doi.org/10.1093/femsre/fuv008
  37. Murzyn K, Rog T, Pasenkiewicz-Gierula M. 2005. Phosphatidylethanolamine-phosphatidylglycerol bilayer as a model of the inner bacterial membrane. Biophys. J. 88: 1091-1103. https://doi.org/10.1529/biophysj.104.048835
  38. Epand RM, Epand RF, Savage PB. 2008. Ceragenins (cationic steroid compounds), a novel class of antimicrobial agents. Drug News Perspect. 21: 307-311. https://doi.org/10.1358/dnp.2008.21.6.1246829
  39. Pichler H, Emmerstorfer-Augustin A. 2018. Modification of membrane lipid compositions in single-celled organisms - From basics to applications. Methods 147: 50-65. https://doi.org/10.1016/j.ymeth.2018.06.009
  40. Gopal R, Seo CH, Song PI, Park Y. 2013. Effect of repetitive lysine-tryptophan motifs on the bactericidal activity of antimicrobial peptides. Amino Acids 44: 645-660. https://doi.org/10.1007/s00726-012-1388-6
  41. Coccia C, Rinaldi AC, Luca V, Barra D, Bozzi A, Di Giulio A, et al. 2011. Membrane interaction and antibacterial properties of two mildly cationic peptide diastereomers, bombinins H2 and H4, isolated from Bombina skin. Eur. Biophys. J. 40: 577-588. https://doi.org/10.1007/s00249-011-0681-8
  42. Lee JU, Park KH, Lee JY, Kim J, Shin SY, Park Y, et al. 2008. Cell selectivity of arenicin-1 and its derivative with two disulfide bonds. Bull. Korean Chem. Soc. 29: 1190-1194. https://doi.org/10.5012/bkcs.2008.29.6.1190
  43. Gotts JE, Matthay MA. 2016. Sepsis: pathophysiology and clinical management. BMJ 353: i1585. https://doi.org/10.1136/bmj.i1585
  44. Ryu DW, Kim HA, Song H, Kim S, Lee M. 2011. Amphiphilic peptides with arginines and valines for the delivery of plasmid DNA. J. Cell. Biochem. 112: 1458-1466. https://doi.org/10.1002/jcb.23064
  45. de Jesus AJ, Allen TW. 2013. The role of tryptophan side chains in membrane protein anchoring and hydrophobic mismatch. Biochim. Biophys. Acta 1828: 864-876. https://doi.org/10.1016/j.bbamem.2012.09.009
  46. Chan DI, Prenner EJ, Vogel HJ. 2006. Tryptophan- and arginine-rich antimicrobial peptides: structures and mechanisms of action. Biochim. Biophys. Acta 1758: 1184-1202. https://doi.org/10.1016/j.bbamem.2006.04.006
  47. Schibli DJ, Epand RF, Vogel HJ, Epand RM. 2002. Tryptophan-rich antimicrobial peptides: comparative properties and membrane interactions. Biochem. Cell Biol. 80: 667-677. https://doi.org/10.1139/o02-147

Cited by

  1. Development of a Novel Short Synthetic Antibacterial Peptide Derived from the Swallowtail Butterfly Papilio xuthus Larvae vol.30, pp.9, 2019, https://doi.org/10.4014/jmb.2003.03009
  2. Anti-Endotoxin 9-Meric Peptide with Therapeutic Potential for the Treatment of Endotoxemia vol.31, pp.1, 2019, https://doi.org/10.4014/jmb.2011.11011