Sains Malaysiana 52(11)(2023): 3223-3237

http://doi.org/10.17576/jsm-2023-5211-15

In silico Design and Evaluation of Novel Cell Targeting Melittin-Interleukin-24 Fusion Protein: A Potential Drug Candidate Against Breast Cancer

(Reka Bentuk in silico dan Penilaian Penyasaran Sel Baru Protein Gabungan Melitin-Interleukin-24: Calon Dadah Berpotensi Melawan Kanser Payudara)

HAFIZ MUHAMMAD REHMAN^1,4, HAFIZ MUZZAMMEL REHMAN^5,6, NADEEM AHMED², MUHAMMAD IMRAN AMIRZADA³, SHAKIRA ASLAM¹ & HAMID BASHIR^1,*

       ¹Centre for Applied Molecular Biology (CAMB), 87-West Canal, Bank Road, University of the Punjab, Lahore-53700, Pakistan

           ²Centre of excellence in Molecular Biology (CEMB), 87-West Canal, Bank Road, University of the Punjab, Lahore-53700, Pakistan

            ³Department of Pharmacy COMSATS University, Islamabad Abbottabad, Pakistan

     ⁴University Institute of Medical Laboratory Technology, Faculty of Allied Health Sciences, The University of Lahore -54590 Pakistan

     ⁵School of Biochemistry and Biotechnology University of the Punjab, Lahore-53700, Pakistan

    ⁶Department of Human Genetics and Molecular Biology, University of Health Science, Lahore

Received: 13 December 2022/Accepted: 16 October 2023

Abstract

Fusion proteins are designed to achieve new functionality or improved properties synergistically by incorporating multiple protein domains into one complex. The fusion of two genes to translate a recombinant protein for cancer treatment can enhance the bioactivity of drug and can introduce novel drug candidate with wide range of applications in pharmaceuticals and biotechnology. Interleukin-24 (IL-24) is a novel cancer growth-suppressing and apoptosis inducing cytokine while melittin is a natural honeybee derived cationic polypeptide having anti-tumor activity against breast cancer cells. The current study was aimed to perform in silico design and analyses of a melittin-IL-24 fusion protein against breast cancer. The amino acid sequences of the IL-24 and melittin peptide were used to design the fusion protein via a rigid linker. Using the online softwares we predicted the secondary and tertiary structures along with physicochemical properties of the designed fusion protein. The validation and quality of the fusion protein was confirmed by Rampage and ERRAT2. The top ranked structure from I-TASSER showed 18.1KD molecular weight by ProtParam, quality factor of 94.152 by ERRAT and a valid structure by Ramachandran plot with 88.5% residues in favoured region. The docking and simulation studies were performed using ClusPro and Desmond software. The quality, validity, interaction analysis and stability of the fusion protein depicted a functional molecule. The in silico analysis finding and expression predicted value of 0.86 in E. coli on SOLUPROT tool suggest that the melittin- IL-24 fusion protein can lead to develop a potent therapeutic drug against breast cancer.

Keywords: Breast cancer; fusion protein; interleukin 24; melittin; molecular docking

Abstrak

Protein gabungan telah direka bentuk untuk mencapai fungsi baharu atau sifat yang dipertingkatkan secara sinergi dengan menggabungkan berbilang domain protein ke dalam satu kompleks. Gabungan dua gen untuk menterjemah protein rekombinan untuk rawatan kanser boleh meningkatkan bioaktiviti dadah dan boleh memperkenalkan calon dadah baharu dengan pelbagai pengaplikasian dalam farmaseutikal dan bioteknologi. Interleukin-24 (IL-24) ialah sitokin penekan pertumbuhan kanser baharu dan apoptosis manakala melitin ialah polipeptida kation perolehan lebah madu semula jadi yang mempunyai aktiviti anti-tumor terhadap sel kanser payudara. Kajian semasa ini bertujuan untuk menjalankan reka bentuk in silico dan analisis protein gabungan melitin-IL-24 terhadap kanser payudara. Jujukan asid amino IL-24 dan peptida melitin digunakan untuk mereka bentuk protein gabungan melalui penghubung tegar. Dengan menggunakan perisian atas talian, kami meramalkan struktur sekunder dan tertiari bersama-sama dengan sifat fizikokimia reka bentuk protein gabungan. Pengesahan dan kualiti protein gabungan telah disahkan oleh Rampage dan ERRAT2. Struktur kedudukan teratas daripada I-TASSER menunjukkan berat molekul 18.1KD oleh ProtParam, faktor kualiti 94.152 oleh ERRAT dan struktur yang sahih oleh plot Ramachandran dengan 88.5% residu di kawasan yang digemari. Kajian dok dan simulasi dilakukan menggunakan perisian ClusPro dan Desmond. Kualiti, kesahan, analisis interaksi dan kestabilan protein gabungan menggambarkan molekul fungsian. Penemuan analisis in silico dan pengekspresan nilai ramalan 0.86 dalam E. coli pada perkakas SOLUPROT menunjukkan bahawa protein gabungan melitin-IL-24 boleh membawa kepada pembangunan ubat terapeutik poten terhadap kanser payudara.

Kata kunci: Dok molekul; interleukin 24; kanser payudara; melitin; protein gabungan

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*Corresponding author; email: hamid.camb@pu.edu.pk