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CxxC Zinc Finger Protein Derived Peptide, MF18 Functions Against Biofilm Formation

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Abstract

The major threat in modern medicine was biofilm forming bacterial related infections and they were highly tolerant to conventional antibiotics and a boundless demand for new drugs. In this regard, antimicrobial peptide (AMP) have been considered as potential alternative agents to conventional antibiotics. In this study, we have reported a CxxC zinc finger protein derived peptide, MF18 and its various biological role including activity against biofilm forming bacteria. Zinc finger protein are important in regulation of several cellular processes and wide range of molecular functions. The CxxC zinc finger protein identified from the cDNA library of a teleost fish; further it was characterised using various online bioinformatics programs. During the in-silico analysis, an AMP named MF18 was identified from the CxxC zinc finger protein, then it was synthesised for further biological activity studies. The antimicrobial activity of MF18 was confirmed against the biofilm clinical isolates such as Staphylococcus aureus and Escherichia coli. The MIC of the antimicrobial peptide at the concentration of 320 µM was observed against these two biofilm bacteria. The mechanism of the peptides was determined using bacteria on its membrane permeabilization ability by scanning electron microscopy. It is exhibited that the MF18 potentially influenced in damaging the morphology of the bacteria. The toxicity of MF18 against the continuous cell line (RAW 264.7) was demonstrated by MTT assay and also using peripheral red blood cells by haemolytic assay; both assays showed that the peptide have no toxicity on the cells at lower concentration. Overall, the study showed the potential therapeutic application of the peptide in pharma industry.

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Abbreviations

ZFP:

Zinc finger protein

PBS:

Phosphate buffered saline

CRA:

Congo red agar

TSB:

Tryptic soy agar

MTT:

3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide

DMEM:

Dulbecco’s modified eagle’s medium

RAW 264.7:

Murine macrophage cell line

OD:

Optical density

DMSO:

Dimethyl sulfoxide

MIC:

Minimum inhibitory concentration

CFU:

Colony forming unit

FACS:

Fluorescent assorted cell sorting

PI:

Propidium iodide

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Authors and Affiliations

  1. Department of Microbiology, SRM Arts and Science College, Kattankulathur, Chennai, 603 203, India

    Prabha Nagaram

  2. Lab PCN 206, Microbiology Division, CSIR-Central Drug Research Institute, Uttar Pradesh, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow, 226 031, India

    Mukesh Pasupuleti

  3. SRM Research Institute, SRM Institute of Science and Technology, Kattankulathur, Chennai, 603 203, India

    Jesu Arockiaraj

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  1. Prabha Nagaram
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Correspondence to Jesu Arockiaraj.

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For human blood collection, informed consent was obtained from all the participated adults in written form as per the Institute ethical approval (Ethical Clearance No. 361/IEC/2012). No animals were involved in this study.

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Nagaram, P., Pasupuleti, M. & Arockiaraj, J. CxxC Zinc Finger Protein Derived Peptide, MF18 Functions Against Biofilm Formation. Protein J 39, 337–349 (2020). https://doi.org/10.1007/s10930-020-09904-1

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