Overexpression of HTRA1 Leads to Ultrastructural Changes in the Elastic Layer of Bruch's Membrane via Cleavage of Extracellular Matrix Components
Figure 6
Degradation of ECM components by recombinant HTRA1.
Purified ECM proteins and purified recombinant HTRA1 were incubated at 37°C in 50 mM Tris-HCl pH7,6, 5 mM CaCl2, 150 mM NaCl over a period of 18 hours (h). (A) Degradation of fibronectin. Fibronectin (FN) shows no sign of degradation when incubated alone. Recombinant HTRA1 degrades fibronection after three hours of incubation. The inhibitor of HTRA1 completely abolishes degradation of fibronectin by HTRA1. (B) Degradation of nidogen 1. Nidogen 1 (NID1) shows no sign of degradation when incubated alone. Recombinant HTRA1 cleaves nidogen-1 after one hours of incubation. The inhibitor of HTRA1 completely abolishes degradation of nidogen 1 by HtrA1. (C) Degradation of nidogen 2. Nidogen 2 (NID2) shows no sign of degradation when incubated alone. Recombinant HTRA1 cleaves nidogen 2 after one hours of incubation. The inhibitor of HTRA1 completely abolishes degradation of nidogen 2 by HTRA1. (D) Degradation of fibulin 5 by HTRA1. Fibulin 5 (FBLN5) is degraded by HTRA1 already after one hour, but shows no sign of degradation when incubated alone. The inhibitor of HTRA1 completely abolishes degradation of fibulin 5 by HTRA1. (E) Degradation of elastin. Elastin (ELN) shows no sign of degradation when incubated alone and when incubated with recombinant HTRA1.