Turbocharged Chemical Protein Synthesis – Turbo

In recent years, chemical protein synthesis enabled the production of homogeneous batches of large and highly decorated protein targets. In doing so, it has provided the means for successful applications in chemical biology, medicinal chemistry, materials science and nanotechnology research.
Chemical protein synthesis primarily relies on the chemoselective coupling of unprotected peptide segments in aqueous media using amide ligation chemistries. The native chemical ligation (NCL) is the most advanced and popular ligation method so far. Nevertheless, the use of NCL for chemical protein synthesis from shorter peptide segments is faced with three major challenges: (1) the diversity of the peptide junctions that has to be made, (2) the modest rate of peptide bond formation and finally (3) the variable solubility of the peptide segments that are linked together.
In this context, we have recently discovered a powerful mean for accelerating chemical reactions in water including NCL by mimicking a mechanism used by cells to promote the association of protein to various biomolecules. In particular, we showed that equipping a thioester with phosphoserines and the Cys partner with an oligoarginine enables outstanding rate accelerations of four order of magnitude for NCL. Although powerful, the NCL process we reported leads to the incorporation of extra charged amino acids in the final product.
With this knowledge, the objective of Turbo project is to develop an innovative electrostatically-assisted and traceless NCL process enabling any type of junction to be formed and extremely fast ligation rates to be achieved. Because the electrostatic assistance relies on the use of oligoarginine modules, we also expect with this approach to significantly improve the solubility of poorly soluble sequences. Therefore, the proposed strategy is ambitious by providing an all-in-one solution to the three challenges discussed above.