Modeling Of Tumor Associated Mature MMP14Protein And The Screening Of The Targeted Peptide Lead For The Divergent Region Of MMP14

Local invasion and distant metastasis are the most important characteristics of malignant tumors, which is the main reason leading to the death of the patient. Matrix metalloproteinases (MMPs) are a class of zinc-dependent endoprotease, playing a key role in cancer development and migration. MMP14(also known as MT1-MMP) is a membrane-type MMP that was overexpressed in many tumor cell surfaces, it can degrade various components of the extracellular matrix as well as activate MMP2and MMP13, being considered as a key enzyme in tumor invasion and migration. MMP14inhibitor has become a hot spot in tumor research and developing a specific inhibitors of MMP14is of great significance in cancer diagnosis and treatment. However, due to the high conservation of the MMPs family and lacking of elucidated crystal structure for the mature MMP14, there is no MMP14or MMPs based anticancer drugs in the market till now. Therefore, the modeling of the three-dimensional structure of mature MMP14and finding of MMP14specific region are significant to design of MMP14specifically binding inhibitors. In this study, the three-dimensional structure of mature MMP14was constructed by homology and de novo modeling and later rationalized and optimized by molecular dynamics simulations. Results of model detection by acknowledged servers,MolProbity, PROCHECK and ERRAT, demonstrated that our models were highly evaluated. An antisense peptide library was constructed against the divergent sense peptide DEGTEEET in the specific region of MMP14which was found by multiple alignment of the whole MMPs family. The antisense peptide library was virtually screened against the constructed three-dimensional model of MMP14by Libdock and further evaluated and optimized by MVD. The top20novel peptides were screened and found well docked with MMP14at the region of DEGTEEET, which indicates they have specific binding ability to MMP14. Preliminary study of one of the top ranked peptides showed that it can inhibit the viabilities of MG-63and MDA-MB-231tumor cells, both of which have expression of MMP14on the cell surface, while it has no obvious effect to MCF-7tumor cells without expression of MMP14, which indicates that it was specific targeting for MMP14. On the other hand,5rounds phage display random peptide library screening were carried out by using the synthesised peptide DEGTEEET as a target.The phages were effectively enriched with the increase in the number of screening rounds and totally10novel dodecapeptide binding to MMP14were obtained.This work not only successfully modeled the three-dimensional structure of mature MMP14but also obtained MMP14target binding peptides by antisense peptide screening and phage random peptide library screening against specific region of MMP14.This work provided an important basis as well as a new way for the discovery and study of MMP14targeted small molecule peptide-based drugs.