The present proposal aims at the characteriazion of the intein-mediated protein splicing
mechanism. Inteins are a widely distributed family of self-splicing proteins, just like RNA splicing, with an ability to excise themselves from flanking host protein regions, the exteins, in an autocatalytic manner with remarkable precision, ligating flanked host protein fragments.
Split inteins are an especially interesting subfamily of inteins. In this case intein is split into two pieces and splicing only
occurs upon reconstitution of these fragments: they generate a single protein chain from two individual polypeptides. Protein-splicing technology is already adapted to a wide range of applications, starting with untagged protein purification, site-specific protein labeling, protein biotinylation, isotope incorporation, peptide cyclization, as antimicrobial target, and so on.
However, structural features of inteins influencing the protein splicing reaction steps, controlling their efficiency and general applicability are poorly understood. We aim at generating a comprehensive analysis of the behaviour of protein splicing reaction/components in order to shedding light on the role played by the split-inteins by means of state-of-the-art spectroscopic and computational approaches.
