Molecular dissection of TIMP3 mutation S156C associated with Sorsby fundus dystrophy
| Autor: | Marton Fogarasi, Andreas Janssen, Bernhard H. F. Weber, Heidi Stöhr |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Vascular Endothelial Growth Factor A
Proteases Angiogenesis Gene Expression Neovascularization Physiologic ADAM17 Protein Matrix metalloproteinase Biology medicine.disease_cause Neovascularization Macular Degeneration Mice Chondrocytes Matrix Metalloproteinase 13 Gene expression medicine Animals Aggrecans Receptor Molecular Biology Cells Cultured Mice Knockout Tissue Inhibitor of Metalloproteinase-3 Tissue Inhibitor of Metalloproteinase-2 Mutation Tissue Inhibitor of Metalloproteinase-1 Endothelial Cells Tissue Inhibitor of Metalloproteinases Kinase insert domain receptor Fibroblasts Vascular Endothelial Growth Factor Receptor-2 Molecular biology Recombinant Proteins ADAM Proteins Amino Acid Substitution Liver ADAMTS4 Protein ADAMTS5 Protein medicine.symptom Procollagen N-Endopeptidase |
| Zdroj: | Matrix Biology. 27:381-392 |
| ISSN: | 0945-053X |
| DOI: | 10.1016/j.matbio.2008.01.008 |
| Popis: | Sorsby fundus dystrophy (SFD) is an autosomal dominant macular degeneration of late onset. A key feature of the disease is the thickening of Bruch's membrane, an ECM structure located between the RPE and the choroid. SFD is caused by mutations in the gene encoding the ECM-associated tissue inhibitor of metalloproteases-3 (TIMP3). We have recently generated two Timp3 gene-targeted mouse lines, one deficient for the murine gene (Timp3-/-) and one carrying an SFD-related S156C mutation. Based on extracts and cell cultures derived from tissues of these animals we now evaluated TIMP3 functionality and its contribution to SFD. We show that the activity levels of TIMP3 target proteases including TACE, ADAMTS4/5 and aggrecan-cleaving MMPs are similar in Timp3S156/+ and Timp3S156C/S156C mice when compared to controls. In Timp3-/- mice, a significant enhancement of enzyme activity was observed for TACE but not for ADAMTS4/5 and MMPs indicating a compensatory effect of other inhibitors regulating the latter two groups of proteases. Fibrin bead assays show that angiogenesis in Timp3S156/+ and Timp3S156C/S156C mice is not altered whereas increased formation of capillary tubes was observed in Timp3-/- animals over controls. Rescue experiments using recombinant proteins demonstrate that the inhibitory activities of TIMP3 towards TACE and aggrecan-cleaving MMPs as well as the anti-angiogenic properties of TIMP3 are not impaired by SFD mutation S156C. We finally demonstrate that wild-type and S156C-TIMP3 proteins block the binding of VEGF to its receptor VEGFR2 to a similar extent. Taken together, this study shows that S156C-TIMP3 retains its known functional properties suggesting that causes other than an imbalance in protease or angiogenic activities represent the primary molecular defect underlying SFD. |
| Databáze: | OpenAIRE |
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