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The Synthetic Antimicrobial Peptide Derived From Melittin Displays Low Toxicity and Anti-infectious Properties

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Abstract

The low stability and nonspecific toxicity are the main limiting factors for the clinical applications of melittin (MLT). This study aimed to design and synthesize new analogs of MLT to increase stability, reduce toxicity, and retain their antimicrobial properties against bacterial pathogens. At first, peptide analogs were designed computationally by inducing single mutations in MLT peptides and evaluating their physicochemical properties. The stability of the analogs with the highest scores was determined by Gromacs software. In vitro assays were performed to examine the antimicrobial activity and toxicity of the selected analogs. Two peptide analogs, M1 and M2, were selected based on the SVM score in cell PPD. The M1 analog was created by replacing alanine with leucine on the 15th. The M2 analog was designed by substituting alanine with leucine and isoleucine with arginine at the 15th and 17th positions. According to the Gromacs results, the M2 peptide indicated more stability. RMSD and RMSF results showed no undesirable fluctuations during the 200 ns MD simulation. The MIC and MBC values for the M1 peptide were calculated in a range of 8–128 μg/ml, while the M2 peptide limited the bacterial growth to 32–128 μg/mL. Both peptides indicated less toxicity than natural MLT, based on MTT assay results. The hemolytic activity of the M1 analog was more than M2 at 16 μg/mL concentration. M1 peptide displayed the highest selectivity index against S. aureus and A. baumannii, which were approximately 5.27-fold improvements compared to MLT. In conclusion, we introduced two analogs of MLT with low toxicity, low hemolytic activity, and higher stability, along with retaining antimicrobial properties against gram-negative and positive bacteria.

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Abbreviations

AMPs:

Antimicrobial peptides

M1:

GLPLLISWIKRKRQQ

M2:

GLPLLRSWIKRKRQQ

GRAVY index:

The sum of the hydropathy values

D:

Discrimination factor

H:

Mean hydrophobicity

μH:

The hydrophobic moment

z:

Net charge

NPT:

Constant pressure

NVT:

Constant volume

RMSF:

Root-mean-square fluctuations

RMSD:

Root-mean-square deviation

DPPC:

Dipalmitoyl phosphatidylcholine membrane

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Acknowledgements

This study was supported by the Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Author information

Authors and Affiliations

  1. Infectious and Tropical Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Parisa Mansouri Rad, Leila Rahbarnia & Zahra Maani

  2. Connective Tissue Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

    Azam Safary

  3. Research Center for Pharmaceutical Nanotechnology, Biomedicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran

    Azam Safary

  4. Plant Biotechnology, Atatürk University, Erzurum, Turkey

    Azizeh ShadiDizaji

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  1. Parisa Mansouri Rad
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  2. Leila Rahbarnia
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  3. Azam Safary
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  4. Azizeh ShadiDizaji
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  5. Zahra Maani
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Contributions

L.R. designed the study and revised the manuscript; A.S. performed the data analysis and interpretation. P.M. and A.Sh wrote the first draft of the manuscript. Z.M. discussed the results. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Leila Rahbarnia.

Ethics declarations

Ethics Approval and Consent to Participate

The study was conducted after ethical approval of the ethics committee of Tabriz University of medical science, Tabriz Iran (reference number: IR.TBZMED.REC.1401.121). Data and clinical samples were collected after written consent with a brief description about the importance of the study to the participants

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The authors declare no competing interests.

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Rad, P.M., Rahbarnia, L., Safary, A. et al. The Synthetic Antimicrobial Peptide Derived From Melittin Displays Low Toxicity and Anti-infectious Properties. Probiotics & Antimicro. Prot. 16, 490–500 (2024). https://doi.org/10.1007/s12602-023-10066-6

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  • DOI: https://doi.org/10.1007/s12602-023-10066-6

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