Expression Of Hemopexin-Like Domain Of Human Matrix Metalloproteinase 2 In E.Coli And It Bioactivity Analysis

Hemopexin-like Domain of human Matrix Human Matrix Metalloproteinase-2 (PEX) is the fragment of C termination of Human matrix Metalloproeteinase- 2(MMP-2). PEX of the dissolvable MMP can band specifically with a kind of cross-membrane protein namedαυβ3 in the membrane and make the MMP rooted on the surface of the cell then to activate the catalyzing domain so as to dissolve cytoplasm and destroy the basement membrane. Under normal physiological condition, MMP supplies necessary room for cell to grow by its dissolving activity, but when tumors or other pathological condition exist, it will express in a high level and help the tumors form and transport by improving the forming of the vein of the tumors. PEX, the fragment of the C termination of MMP, can interrupt the banding between MMP andαυβ3 by competitive banding with the latter and protect the plasm between cells from being destroyed, by all these, to restrain the forming of new vein. Basing on a method of curing the tumor called dormancy therapy, PEX of MMP-2 is always being considered as a hopeful candidate medicament in restraining the forming of the tumor vein. Now, PEX of MMP-2 has been expressed in prokaryote and eukaryote according to the foreign reports, and the product could remarkably restrain the growth and transportation of tumor tissue by repressing the forming of new vein; there is also report about expressing PEX of MMP-2 in prokaryote and restraining the forming of new vein.Using the vector of expressing PEX of MMP-2 in prokaryote being construct in our lab, to induce the PEX expressing, as while, considering the product cost, a 40% expression of PEX combination protein has got by optimizing the concentration of IPTG, OD value, inducing time and PH value of the culture medium. Comparing three plan for abstergent PEX inclusion bodies, complex abstergent could got purer inclusion body protein and the amount of inclusion body protein after abstergent, analyzing by software, could got 86% of the total protein; here we also have three different renaturation methods for the inclusion body respectively dissolved in urea and Guanidine HCl and calculate the renaturation ratio, and by chicken CAMs, the renaturation product has the activity of restraining the forming of new vein. Through choosing simple and economical renaturation methods, this study did a good try on speeding up the commercial production of recombined PEX protein.The main results we got in this study are:...