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 AHMED 2 , MUHAMMAD IMRAN AMIRZADA 3 , SHAKIRA ASLAM 1 & HAMID BASHIR 1,* 1 Centre for Applied Molecular Biology (CAMB), 87-West Canal, Bank Road, University of the Punjab, Lahore-53700, Pakistan 2 Centre of excellence in Molecular Biology (CEMB), 87-West Canal, Bank Road, University of the Punjab, Lahore-53700, Pakistan 3 Department of Pharmacy COMSATS University, Islamabad Abbottabad, Pakistan 4 University Institute of Medical Laboratory Technology, Faculty of Allied Health Sciences, The University of Lahore -54590 Pakistan 5 School of Biochemistry and Biotechnology University of the Punjab, Lahore-53700, Pakistan 6 Department 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