In silico directed evolution of Anabas testudineus AtMP1 antimicrobial peptide to improve in vitro anticancer activity

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Biochemistry, Biophysics and Molecular Biology

Main article text

 

Introduction

Materials & Methods

Antimicrobial peptide design and alteration

Prediction of peptide properties using bioinformatic tools

ANTICP database

Prot param tool

Prediction of the toxicity of selected peptides

ADME studies

Allergenicity prediction

Synthesis of the AMPS

Cytotoxicity effect of the peptide

Cell lines

Anticancer activity of peptides

Determination of cell apoptosis

Annexin V-FITC apoptosis

Cell cycle profile

RNA isolation and cDNA synthesis

Data analysis

Results & Discussion

Results of peptide alteration

Selection of the potential AMPs

Results of (ADME) prediction

Results of cytotoxic effect of AMPs

Apoptosis induction by annexin V-FTIC assay

Cell cycle analysis

Gene expression profiler RT2 PCR array results

Conclusions

Supplemental Information

Raw cytotoxicity data.

DOI: 10.7717/peerj.17894/supp-1

The complete list of modified peptides and their predicted properties using in silico databases (ADP3, AMPfun, and CAMP R3)

DOI: 10.7717/peerj.17894/supp-2

The complete results of the ADME in silico prediction a multitude of crucial parameters were evaluated, encompassing the partition coefficient (log P), the permeability of Madin Darby canine kidney cells (MDCK), the capacity to function as a P-glycoprotei

DOI: 10.7717/peerj.17894/supp-3

Additional Information and Declarations

Competing Interests

The authors declare there are no competing interests except Choy-Theng Loh is employed by Hangzhou Foreseebio Biotechnology Co., Ltd and being appointed as adjunct Faculty Member at INTI International University. All the work done by was Choy-Theng Loh was purely academic based.

Author Contributions

Shazrul Fazry conceived and designed the experiments, prepared figures and/or tables, and approved the final draft.

Ahmed Abdulkareem Najm conceived and designed the experiments, performed the experiments, prepared figures and/or tables, and approved the final draft.

Ibrahim Mahmood Mahdi performed the experiments, authored or reviewed drafts of the article, and approved the final draft.

Arnold Ang performed the experiments, prepared figures and/or tables, and approved the final draft.

LiTing Lee analyzed the data, prepared figures and/or tables, and approved the final draft.

Choy-Theng Loh analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Sharifah Sakinah Syed Alwi analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Fang Li analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

Douglas Law conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Data Availability

The following information was supplied regarding data availability:

Raw data are available in the Supplemental Files.

Funding

This study was supported by the Ministry of Higher Education (MoHE), Malaysia (FRGS/1/2022/STG01/INTI/02/3) & (FRGS/1/2023/STG01/UKM/02/5), Universiti Kebangsaan Malaysia (ST-2022-013). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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