Converting a broad matrix metalloproteinase family inhibitor into a specific inhibitor of MMP MMP ‐9 and MMP MMP ‐14

摘要

MMP ‐14 and MMP ‐9 are two well‐established cancer targets for which no specific clinically relevant inhibitor is available. Using a powerful combination of computational design and yeast surface display technology, we engineered such an inhibitor starting from a nonspecific MMP inhibitor, N‐ TIMP 2. The engineered purified N‐ TIMP 2 variants showed enhanced specificity toward MMP ‐14 and MMP ‐9 relative to a panel of off‐target MMP s. MMP ‐specific N‐ TIMP 2 sequence signatures were obtained that could be understood from the structural perspective of MMP /N‐ TIMP 2 interactions. Our MMP ‐9 inhibitor exhibited 1000‐fold preference for MMP ‐9 vs. MMP ‐14, which is likely to translate into significant differences under physiological conditions. Our results provide new insights regarding evolution of promiscuous proteins and optimization strategies for design of inhibitors with single‐target specificities.