Improving the Stability of Anterior Gradient ۲ Protein Using Rational Design

Despite major advances in wound healing, healing remains one of the major challenges ahead. One of the effective factors in improving the wound healing process is AGR۲ protein and can be effective as a local factor in the healing process. Therefore, increasing thermodynamic stability and ensuring the effectiveness of this protein as a drug is important. To date, no studies have been performed to increase the thermodynamic stability of this protein using point mutations. The aim of this study was to improve the thermodynamic stability of a protein using targeted point mutations in its amino acid sequence. For this purpose, the structure of AGR۲ protein in the PDB database was first systematically analyzed using Dezyme software and the parts of the structure that are most likely to be stable after mutation were identified. By comparing these data with information from previous structural studies on protected and unprotected areas, a total of ۳ unprotected points were selected to create point mutations. Using the DynaMut server, we first predicted changes in protein stability due to Y۱۵۰K, N۱۶۳F, and S۶۵L point mutations individually. Then, in a study, the effect of all ۳ mutations together on the stability of the protein by the mentioned server was studied. The results of these studies showed that the stabilizing effect of Y۱۵۰K, N۱۶۳F and S۶۵L mutations were ۰.۳۷۴, ۰.۵۳۳ and ۲.۱۶۵ kcal/mol, respectively. Also, the stabilizing effect of these three mutations together on the stability of the protein was approximately ۱Kcal / mol. Therefore, applying all three of these mutations together has a better effect on increasing the thermodynamic stability of the protein by reducing energy. Experimental confirmation of the effect of these mutations on protein stability and their in vitro studies could offer new perspectives in this field.

کلیدواژه ها:

Site-directed muttion ، Wound healing ، Dezyme Software ، DynaMut server