Anti-TIMP2 (full length) polyclonal antibody

OBJECTIVE: To explore the correlations of the tissue inhibitor of metalloproteinase-2 (TIMP2) and TIMP3 gene polymorphisms with primary open-angle glaucoma (POAG). PATIENTS AND METHODS: 295 POAG patients were recruited as the observation group and 200 healthy people as the control group. The general clinical information, medical history (hypertension, diabetes and hyperlipemia) and family history were collected. The vascular endothelium and hemorheology indexes in each group were detected. Moreover, the polymorphisms of TIMP2 rs796391657 and TIMP3 rs8136803 were detected via TaqMan-MGB probe assay. RESULTS: The observation group exhibited higher prevalence rates of diabetes, hyperlipemia and family history than those of the control group (p<0.05), and there were no differences in age, sex and hypertension prevalence rate between the two groups (p>0.05). The observation group had a lower nitric oxide (NO) level, but higher levels of endothelin-1 (ET-1), plasma viscosity (PV), hematocrit (HCT) and fibrinogen (FIB) than those of the control group (p<0.05). The distribution of the three genotypes and alleles of TIMP2 rs796391657 varied a lot between the two groups (p<0.05). The frequency of genotype CC was markedly higher than that of genotype TT; however, C allele had a higher frequency than T allele. Similarly, the three genotypes and alleles of TIMP3 rs8136803 were differentially distributed in the two groups (p<0.05). The frequency of genotype GG was higher than that of genotype TT, while the frequency of G allele was higher than that of T allele. CONCLUSIONS: The TIMP2 and TIMP3 gene polymorphisms are correlated with POAG.

Degradation of the extracellular matrices in the human body is controlled by matrix metalloproteinases (MMPs), a family of more than 20 homologous enzymes. Imbalance in MMP activity can result in many diseases, such as arthritis, cardiovascular diseases, neurological disorders, fibrosis, and cancers. Thus, MMPs present attractive targets for drug design and have been a focus for inhibitor design for as long as 3 decades. Yet, to date, all MMP inhibitors have failed in clinical trials because of their broad activity against numerous MMP family members and the serious side effects of the proposed treatment. In this study, we integrated a computational method and a yeast surface display technique to obtain highly specific inhibitors of MMP-14 by modifying the natural non-specific broad MMP inhibitor protein N-TIMP2 to interact optimally with MMP-14. We identified an N-TIMP2 mutant, with five mutations in its interface, that has an MMP-14 inhibition constant (K-i) of 0.9 pm, the strongest MMP-14 inhibitor reported so far. Compared with wild-type N-TIMP2, this variant displays approximate to 900-fold improved affinity toward MMP-14 and up to 16,000-fold greater specificity toward MMP-14 relative to other MMPs. In an in vitro and cell-based model of MMP-dependent breast cancer cellular invasiveness, this N-TIMP2 mutant acted as a functional inhibitor. Thus, our study demonstrates the enormous potential of a combined computational/directed evolution approach to protein engineering. Furthermore, it offers fundamental clues into the molecular basis of MMP regulation by N-TIMP2 and identifies a promising MMP-14 inhibitor as a starting point for the development of protein-based anticancer therapeutics.