Method of protein stabilization based on prediction of intrinsically disordered regions. Stabilization of protein Gαo

Abstract

Protein Gαo from Drosophila melanogaster has been studied. We have designed an SS-bridge in this protein in such a way that its stability could be increased. The choice of a region in the polypeptide chain for insertion of the cysteine bridge was based on calculations performed using programs for the prediction of intrinsically disordered proteins. In our previous paper we proposed that, in structured proteins, these programs predict “weakened” protein regions unable to have a rigid structure without additional hydrophobic interactions rather than intrinsically disordered regions. Consequently, insertion of SS-bridges in such regions will most probably lead to stabilization of the protein. It was just this approach that was used in the presented study. It is demonstrated with the microcalorimetry method that the insertion of the designed SS-bridge resulted in 4 K increase in the melting temperature of one domain of protein Gαo. Refs 20. Figs 3. Tables 1.