Computational design and evaluation of a novel temporin 1CEa-IL24 fusion protein for anti-tumor potential

Introduction: Synthetic fusion proteins represent a cutting-edge approach in biotechnology and pharmaceutical research, enabling the strategic combination of multiple protein domains to design novel complexes with enhanced properties and functionalities. By fusing distinct proteins, the unique attributes of each component can be synergistically exploited, leading to improved bioactivities or the emergence of entirely new functions. This study aimed to computationally construct a fusion protein that contained the killing properties of temporin 1CEa and the targeting action of IL-24, joined via a rigid linker, and its anti-tumor potential was analyzed using different bioinformatics tools. Methods: After sequence retrieval, the 3D model of the temporin 1CEa-IL-24 fusion protein was constructed using the AlphaFold 2 online server. The resulting structure underwent rigorous refinement, quality assessment, and validation processes. Physiochemical properties were evaluated using ProtParam, and the prepared structure was subjected to docking on ClusPro and simulation on VMD and NAMD. Results: Assessment through ERRAT score and Ramachandran plot analysis confirmed the good quality, refinement, and validation of the 3D structure of the temporin 1CEa-IL-24 fusion protein. Docking results revealed 17 hydrogen bonds and 4 salt bridges between the fusion protein and its receptor, findings supported and validated through simulation studies predicting a stable docked complex over a 100 ns period. Conclusion: The expression of the temporin 1CEa-IL-24 fusion gene in a suitable expression host could lead to in vitro production and subsequently validate its therapeutic potential.