Figure 3. Purification of N-TIMP2 selective variants.
(A) SEC (using Superdex75) analysis of clone N-TIMP214_17. (B) Mass spectrometry analysis of clone N-TIMP214_17 after SEC. (C) Analysis of purified N-TIMP2WT, N-TIMP214_17, N-TIMP214_18, N-TIMP29_1 and N-TIMP29_13 by 15% SDS-PAGE performed under reducing conditions. (D) MMP14CAT inhibition by various concentrations of N-TIMP2WT, N-TIMP214_17 and N-TMP214_18. Cleavage of the fluorescent substrate was measured over time, with the velocity (slope) of the reaction as a function of inhibitor concentration being fitted to Morrison's equation to obtain the inhibition constant Ki. (E) MMP14CAT inhibition by various concentrations of N-TIMP29_1 and N-TIMP29_13. (F) Inhibition of MMP9CAT by N-TIMP2WT and selective inhibitors. (G) Inhibition of MMP1CAT by N-TIMP2WT and the selective inhibitors. (H) Inhibition of MMP10CAT by N-TIMP2WT and the selective inhibitors.