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Design and Synthesis of Lipopolysaccharide-Binding Antimicrobial Peptides Based on Truncated Rabbit and Human CAP18 Peptides and Evaluation of Their Action Mechanism

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Abstract

Lipopolysaccharide (LPS) is a toxic and immunogenic agent for human. Additionally, LPS is a good target for some antimicrobial compounds, including antimicrobial peptides (AMPs). LPS-binding peptides (LBPs) can recognize and neutralize LPS. Rabbit and human cathelicidins are AMPs with LPS-binding activity. In this study, we designed and synthesized two new truncated LBPs from rabbit and human CAP18 peptides by in silico methods. After synthesis of peptides, the antimicrobial properties and LPS-binding activity of these peptides were evaluated. The parental rabbit and human CAP18 peptides were selected as positive controls. Next, the changes in the secondary structure of these peptides before and after treatment with LPS were measured by circular dichroism (CD). Human cytotoxicity of the peptides was evaluated by MTT and red blood cells (RBCs) hemolysis assays. Finally, field emission scanning electron microscopy (FE-SEM), confocal microscopy, and flow cytometry were performed to study the action mechanism of these peptides. Results indicated that the hCap18 and rCap18 had antibacterial activity (at a MIC of 4–128 μg/mL). The results of the quantitative LAL test demonstrated that LPS-binding activity of hCap18 peptide was better than rCap18, while rCap18 peptide had better antimicrobial properties. Furthermore, rCap18 had less cytotoxicity than hCap18. However, both peptides were nontoxic for normal human skin fibroblast cell in MIC range. In conclusion, rCap18 has good antibacterial properties, while hCap18 can be tested as a diagnostic molecule in our future studies.

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References

  1. Van Amersfoort ES, Van Berkel TJ, Kuiper J (2003) Receptors, mediators, and mechanisms involved in bacterial sepsis and septic shock. Clin Microbiol Rev 16:379–414

    Article  Google Scholar 

  2. Youn JH, Kwak MS, Wu J, Kim ES, Ji Y, Min HJ, Yoo JH, Choi JE, Cho HS, Shin JS (2011) Identification of lipopolysaccharide-binding peptide regions within HMGB1 and their effects on subclinical endotoxemia in a mouse model. Eur J Immunol 41:2753–2762

    Article  CAS  Google Scholar 

  3. Steimle AIB, Frick JS (2016) Structure and function: lipid a modifications in commensals and pathogens. Int J Med Microbiol 306:290–301

    Article  CAS  Google Scholar 

  4. Park CY, Jung SH, Bak JP, Lee SS, Rhee DK (2005) Comparison of the rabbit pyrogen test and limulus amoebocyte lysate (LAL) assay for endotoxin in hepatitis B vaccines and the effect of aluminum hydroxide. Biologicals 33:145–151

    Article  CAS  Google Scholar 

  5. Raetz CR, Whitfield C (2002) Lipopolysaccharide endotoxins. Annu Rev Biochem 71:635–700

    Article  CAS  Google Scholar 

  6. Novitsky TJ, Roslansky PF, Siber GR, Warren HS (1985) Turbidimetric method for quantifying serum inhibition of limulus amoebocyte lysate. J Clin Microbiol 21:211–216

    Article  CAS  Google Scholar 

  7. Faraj TA, McLaughlin CL, Erridge C (2017) Host defenses against metabolic endotoxaemia and their impact on lipopolysaccharide detection. Int Rev Immunol 36:125–144

    Article  CAS  Google Scholar 

  8. Peters BM, Shirtliff ME, Jabra-Rizk MA (2010) Antimicrobial peptides: primeval molecules or future drugs? PLoS Pathog 6:1–4

    Article  Google Scholar 

  9. Nagaoka I, Hirota S, Niyonsaba F, Hirata M, Adachi Y, Tamura H et al (2002) Augmentation of the lipopolysaccharide-neutralizing activities of human cathelicidin CAP18/LL-37-derived antimicrobial peptides by replacement with hydrophobic and cationic amino acid residues. Clin Diagn Lab Immunol 9:972–982

    CAS  Google Scholar 

  10. Kościuczuk EM, Lisowski P, Jarczak J, Strzałkowska N, Jóźwik A, Horbańczuk J, Bagnicka E et al (2012) Cathelicidins: family of antimicrobial peptides. a review. Mol Biol Rep 39:10957–10970

    Article  Google Scholar 

  11. Larrick JW, Hirata M, Zheng H, Zhong J, Bolin D, Cavaillon JM et al (1994) A novel granulocyte-derived peptide with lipopolysaccharide-neutralizing activity. J Immunol 152:231–240

    CAS  Google Scholar 

  12. Mason DJ, Dybowski R, Larrick JW, Gant VA (1997) Antimicrobial action of rabbit leukocyte CAP18 (106-137). Antimicrob Agents Chemother 41:624–629

    Article  CAS  Google Scholar 

  13. Gutsmann T, Fix M, Larrick JW, Wiese A (2000) Mechanisms of action of rabbit CAP18 on monolayers and liposomes made from endotoxins or phospholipids. J Membr Biol 176:223–236

    Article  CAS  Google Scholar 

  14. Singh H, Srivastava HK, Raghava GP (2016) A web server for analysis, comparison and prediction of protein ligand binding sites. Biol Direct 11:1–14

    Article  Google Scholar 

  15. Balouiri M, Sadiki M, Ibnsouda SK (2016) Methods for in vitro evaluating antimicrobial activity: A review. J Pharm Anal 6:71–79

    Article  Google Scholar 

  16. Manzini MC, Perez KR, Riske KA, Bozelli Jr JC, Santos TL, da Silva MA, Saraiva GK, Politi MJ, Valente AP, Almeida FC, Chaimovich H (2014). Peptide: lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies. Biochim Biophys Acta Biomembr 1838: 1985–1999

  17. Wu X, Wang Z, Li X, Fan Y, He G, Wan Y, Yu C, Tang J, Li M, Zhang X, Zhang H (2014) In vitro and in vivo activities of antimicrobial peptides developed using an amino acid-based activity prediction method. Antimicrob Agents Chemother 58:5342–5349

    Article  Google Scholar 

  18. Madanchi H, Akbari S, Shabani AA, Sardari S, Farmahini Farahani Y, Ghavami G, Ebrahimi Kiasari R (2019) Alignment-based design and synthesis of new antimicrobial aurein-derived peptides with improved activity against Gram-negative bacteria and evaluation of their toxicity on human cells. Drug Dev Res 80:162–170

    Article  CAS  Google Scholar 

  19. Rothan HA, Mohamed Z, Suhaeb AM, Rahman NA, Yusof R (2013) Antiviral cationic peptides as a strategy for innovation in global health therapeutics for dengue virus: high yield production of the biologically active recombinant plectasin peptide. OMICS 17:560–567

    Article  CAS  Google Scholar 

  20. Ghosh A, Datta A, Jana J, Kar RK, Chatterjee C, Chatterjee S, Bhunia A (2014) Sequence context induced antimicrobial activity: insight into lipopolysaccharide permeabilization. Mol BioSyst 10:1596–1612

    Article  CAS  Google Scholar 

  21. Madanchi H, Khalaj V, Jang S, Shabani AA, Ebrahimi Kiasari R, Seyed Mousavi SJ, Sardari S (2019) AurH1: a new heptapeptide derived from aurein1.2 antimicrobial peptide with specific and exclusive fungicidal activity. J Pept Sci 25:1–9

    Article  Google Scholar 

  22. Memariani H, Shahbazzadeh D, Sabatier JM, Memariani M, Karbalaeimahdi A, Bagheri KP (2016) Mechanism of action and in vitro activity of short hybrid antimicrobial peptide PV3 against Pseudomonas aeruginosa. Biochem Biophys Res Commun 479:103–108

    Article  CAS  Google Scholar 

  23. Wiedemann C, Bellstedt P, Görlach M (2013) CAPITO - a web server-based analysis and plotting tool for circular dichroism data. Bioinformatics 29:1750–1757

    Article  CAS  Google Scholar 

  24. Xie P, Zhu L, Shao X, Huang K, Tian J, Xu W (2016) Highly sensitive detection of lipopolysaccharides using an aptasensor based on hybridization chain reaction. Sci Rep 6:1–8

    Article  CAS  Google Scholar 

  25. Cauwels A, Rogge E, Janssen B, Brouckaert P (2010) Reactive oxygen species and small-conductance calcium-dependent potassium channels are key mediators of inflammation-induced hypotension and shock. J Mol Med 88:921–930

    Article  CAS  Google Scholar 

  26. Pedersen MR, Hansen EW, Christensen JD (1994) Detection of lipopolysaccharide in the picogram range of tissue culture media by a kinetic chromogenic limulus amebocyte lysate assay. J Clin Pharm Ther 19:189–194

    Article  CAS  Google Scholar 

  27. Yao M, Zhang H, Dong S, Zhen S, Chen X (2009) Comparison of electrostatic collection and liquid impinging methods when collecting airborne house dust allergens, endotoxin and (1, 3)-β-d-glucans. J Clin Pharm Ther 40:492–502

    CAS  Google Scholar 

  28. Wang Z, Wang X, Wang J (2018) Recent advances in antibacterial and antiendotoxic peptides or proteins from marine resources. Mar Drugs 16:1–18

    Article  Google Scholar 

  29. Maccari G, Nifosi R, Di Luca M (2013) Rational development of antimicrobial peptides for therapeutic use: design and production of highly active compounds. Microbial pathogens and strategies for combating them: science, technology and education 1265-1277

  30. Boland MP, Separovic F (2006) Membrane interactions of antimicrobial peptides from Australian tree frogs. Biochim Biophys Acta Biomembr 1758:1178–1183

    Article  CAS  Google Scholar 

  31. Mor A, Hani K, Nicolas P (1994) The vertebrate peptide antibiotics dermaseptins have overlapping structural features but target specific microorganisms. J Biol Chem 269:31635–31641

    CAS  Google Scholar 

  32. Larrick JW, Hirata M, Zhong J, Wright SC (1995) Anti-microbial activity of human CAP18 peptides. Immunotechnology 1:65–72

    Article  CAS  Google Scholar 

  33. Kaconis Y, Kowalski I, Howe J, Brauser A, Richter W, Razquin-Olazarán I, Iñigo-Pestaña M, Garidel P, Rössle M, Martinez de Tejada G, Gutsmann T, Brandenburg K (2011) Biophysical mechanisms of endotoxin neutralization by cationic amphiphilic peptides. Biophys J 100:2652–2661

    Article  CAS  Google Scholar 

  34. Gutsmann T, Razquin-Olazarán I, Kowalski I, Kaconis Y, Howe J, Bartels R, Hornef M, Schürholz T, Rössle M, Sanchez-Gómez S, Moriyon I, Martinez de Tejada G, Brandenburg K (2010) New antiseptic peptides to protect against endotoxin-mediated shock. Antimicrob Agents Chemother 54:3817–3824

    Article  CAS  Google Scholar 

  35. Heinbockel L, Sánchez-Gómez S, de Tejada GM, Dömming S, Brandenburg J, Kaconis Y et al (2013) Preclinical investigations reveal the broad-spectrum neutralizing activity of peptide Pep19-2.5 on bacterial pathogenicity factors. Antimicrob Agents Chemother 57:1480–1487

    Article  CAS  Google Scholar 

  36. Soufian S, Naderi-Manesh H, Alizadeh A, Sarbolouki M (2009) Molecular dynamics and circular dichroism studies on aurein 1.2 and retro analog. World Acad Sci Eng Tech 56:858–864

    Google Scholar 

Download references

Acknowledgments

We would like to thank the staff of the School of Medicine, Department and Center for Biotechnology Research from Semnan University of Medical Sciences, and Drug Design and Bioinformatics Unit of Pasteur Institute of Iran.

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Author notes

  1. Hamid Madanchi and Ramin Ebrahimi Kiasari are co-first authors and contributed equally to this research

Authors and Affiliations

  1. Department of Biotechnology, School of Medicine, Semnan University of Medical Sciences, Semnan, Iran

    Hamid Madanchi & Ali Akbar Shabani

  2. Drug Design and Bioinformatics Unit, Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

    Hamid Madanchi, Ramin Ebrahimi Kiasari, Seyed Javad Seyed Mousavi & Soroush Sardari

  3. Department of Medical Biotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran

    Behrooz Johari

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  1. Hamid Madanchi
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Correspondence to Soroush Sardari.

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Madanchi, H., Ebrahimi Kiasari, R., Seyed Mousavi, S.J. et al. Design and Synthesis of Lipopolysaccharide-Binding Antimicrobial Peptides Based on Truncated Rabbit and Human CAP18 Peptides and Evaluation of Their Action Mechanism. Probiotics & Antimicro. Prot. 12, 1582–1593 (2020). https://doi.org/10.1007/s12602-020-09648-5

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