The project is based on the very recent development of a set of ferritin protein mutants endowed with unique properties. In particular a genetically engineered construct has been obtained which corresponds to a truncated Ferritin H-homoplymer (¿Cter-HFt) that lacks 13 aminoacids in the C-terminal region and bears selected mutations in order to abolish cysteine residues in the native positions and introduces a new cysteine in a tolopogically selected position for covalent attachment of bioactive small molecule pendants. ¿Cter-HFt possesses the remarkable capabilities in keeping with ferrooxidase activity while it is not capable of iron sequestering and storage. Thus, in the presence of ferrous iron and oxygen in solution, the protein was demonstrated to catalytically generate a high rate of oxygen radicalic species in vitro. The protein has been fully characterized by CryoEM microscopy in order to demonstrate full and correct 24-mer assembly and tested for cell uptake in ligand tracer measurements. Based on these preliminary observations, the present project aims at demonstrating radical species induced cytotoxicity in a number of selected tumor cell lines (overexpressing transferrin receptor), investigate the oxidative damage on these cells and address at least one animal tumor model for possible tumor cytotoxicity effects.
