Design and evaluation of two novel chimeric proteins, p28-IL-24 and p28-M4, targeted to breast cancer cells: An In-Silico approach

Introduction & objectives: p28 is an anticancer peptide (ACP) shown to be cytotoxic against various cancer cells. Moreover, as a cell penetrating peptide (CPP), p28 can be applied as a targeting moiety in the structure of chimeric proteins. IL-24 (or its truncated sequence, M4) is a cytokine with anticancer activity against a wide range of malignant cells. We aimed at production of chimeric proteins composed of P28 and either IL-24 or M4 to target breast cancer cells. However, selection of a suitable linker to join the two moieties is an important factor in the design of chimeric proteins. In the current study, the effect of different linkers on construction of the two chimeric proteins (p28-IL-24 and p28-M4) were evaluated in silico. Materials & Methods: After selection of some linkers with different characteristics, a small library of the chimeric proteins were created and assessed by various online servers and bioinformatics software. Furthermore, following selection of the most appropriate linker, the three dimensional structures and dynamic behavior of both chimeric proteins were evaluated by homology modeling and molecular dynamic simulation (MD), respectively. Results: Based on our results, a rigid linker with AEAAAKEAAAKA sequence showed to provide the highest freedom of action for the both of proteins. Furthermore, according to MD parameters, between p28-IL-24 and p28-M4 chimeric proteins, the first one have better solubility as well as stability and might show stronger anticancer effects in vitro and in vivo, because its moieties showed to exert their activities more freely. Conclusion: Taken together our fining showed that p28-AEAAAKEAAAKA-IL-24 has better stability and solubility than the p28-M4 protein. Therefore this chimeric protein was anticipated to show better anti-cancer efficacy. However in vitro and in vivo studies are needed to assess their biological activity and cytotoxic effects. These studies are undergoing for the two chimeric proteins.

کلیدواژه ها:

p28 ، IL-24 ، M4 ، Linker ، Chimeric protein ، Homology modeling ، Molecular Dynamic Simulation ، Breast cancer