Tissue Factor Pathway Inhibitor 2 (TFPI2) takes part in the regulation of plasmin-mediated matrix remodeling. Inhibits trypsin, plasmin, factor VIIa/tissue factor and weakly factor Xa. TFPI2 doesn’t have any influence on thrombin. TFPI2 also can inhibit MMP activity, which can hydrolyze gelatin under certain conditions. Thus, the activity of TFPI2 can be measured by inhibit MMP-2 hydrolyze gelatin. Gelatin zymography is mainly used for the detection of the gelatinases, 2μg/mL was denatured by SDS loading buffer, electrophoresed through sodium dodecylsulphate–polyacrylamide gel (SDS–PAGE; 8% gels) containing gelatin (1mg/mL) with nonreducing conditions. After renaturation, incubate with various concentrations of recombinant mouse TFPI2, then staining with coomassie brilliant blue G250, active MMP-2 would hydrolyze gelatin nearby, which was indicated by the white binds on the gel; if the activity of MMP-2 inhibit by TFPI2, there was none white binds on the gel.The result was shown in figure 1.
As the figure 1 shown, MMP-2 can be inhibited by recombinant mouse TFPI2 at least 2.5μg/mL.