Degradation of human plasma and extracellular matrix fibronectin by tissue type plasminogen activator and urokinase.
Fibronectins and plasminogen activators, both tissue and urokinase types, are involved in the physiopathological degradation of the extracellular matrix. Previous reports indicate that fibronectin can be degraded by urokinase without plasminogen. Also, we have shown that tissue-type plasminogen activator can cleave fibronectin, without plasminogen, generating fragments of 30 and 220-250 kDa detectable by immunoblotting analysis. A comparison with urokinase-induced degradation indicates that the cleavage sites are the same for both plasminogen activators. One is close to the carboxyl-terminal, disrupting the fibronectin dimeric structure, and one is near the amino-terminal, generating a 30 kDa fragment. In solution, the activity of tissue-type plasminogen activator was prevalent on the amino-terminal site, while urokinase activity was prevalent on the carboxyl-terminal site. On fibronectin immobilized onto a gelatin coated surface, only the 30 kDa fragment was released when treated with both plasminogen activators. Plasminogen activators also were active on fibronectin assembled into the extracellular matrix of cultured fibroblasts. Urokinase caused the complete disappearance of extracellular matrix fibronectin, together with the release of the 30 and the 220-250 kDa fibronectin fragments, but left a flat morphology, while tissue-type plasminogen activator induced the release of the 30 kDa fragment associated with changes in cellular morphology. The plasminogen-independent fibronectin degradation exerted by tissue-type plasminogen activator and urokinase is 100 times lower than that exerted by plasmin. This may provide a mechanism for localized and limited degradation of fibronectin preventing the generalized proteolysis associated with plasminogen activation.
Marchina
E
E
Department of Biomedical Sciences and Biotechnology, University of Brescia.
Barlati
S
S
D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D010959:Tissue Plasminogen Activator / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D005353:Fibronectins; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator