Study of deep eutectic solvent effect on matrixmetalloproteinase-9 stability and structure with treatment purpose

Introduction: Matrix metalloproteinases-9 is a family of zinc dependent endopeptidases commonly known for their abilities to cleave components of the connective tissue such as collagen, elastin and proteoglycan. MMP-9 plays an important role in the development of many diseases such as periodontitis, atherosclerosis and cancer. Therefore, increasing the stability of the enzyme in order to study three-dimensional structure to design of inhibitors with the goal of therapy is very effective. Materials and methods: One of the newly developed methods for stability of enzyme is using Deep Eutectic Solvents (DESs). Herein, active full length recombinant human MMP-9 (amino acid residues 107-707) has been expressed in Escherichia coli BL21, using the vector pET21a. Purification and refolding were conducted using urea gradient method on Ni-NTA column, simultaneously. In this study, the effect of DES based on mixtures of choline chloride and glycerol with a 1:1 mol ratio was investigated on MMP-9 activity, stability and structure. Results: MMp-9 has high activity in presence of 30% v/v DES. The structure of the enzyme by intrinsic fluorescence studies confirmed these results. Compared to buffer, the thermal stability of the enzyme and the remaining activity increased in presence of solvents. Conclusion: Improvement of matrix metalloproteinase-9 thermal stability can be attributed to compactness of structure in the presence of DES in order to evaluate the three-dimensional structure and synthesis of inhibitors with aim of therapy has an enormous effective.

کلیدواژه ها:

Matrixmetalloproteinase-9 (MMP-9) ، Deep eutectic solvent ، Thermal stability ، Remaining activity