Extracellular matrix produced by cultured corneal and aortic endothelial cells contains active tissue-type and urokinase-type plasminogen activators.
Incubation of plasminogen with the subendothelial extracellular matrix (ECM) synthesized by cultured bovine corneal and aortic endothelial cells resulted in generation of fibrinolytic activity, indicated by proteolysis of 125I-fibrin in a time- and dose-dependent manner. Both tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA) were identified in the ECM by fibrin zymography, immunoblotting, and inhibition of plasminogen activation by anti-u-PA and anti-t-PA antibodies. Most of the ECM-resident plasminogen activator (PA) activity did not originate from intracellular PA release occurring when the endothelial cells were lyzed and the ECM exposed, since a comparable amount of PA was associated with the ECM when the cells were lyzed with Triton X-100 or removed intact by treatment with 2 M urea. Active u-PA and t-PA were released from ECM by treatment with heparanase (endo-beta-D-glucuronidase), indicating that some of the ECM-resident PA activity is sequestered by heparan sulfate side chains. These results indicate that both u-PA and t-PA produced by endothelial cells are firmly sequestered in an active form by the subendothelial ECM. It is suggested that ECM-resident plasminogen activators participate in sequential matrix degradation during cell invasion and tumor metastasis. PA activity may also function in release of ECM-bound growth factors (i.e., basic fibroblast growth factor) and activation of proenzymes (i.e., prothrombin), resulting in modulation of the ECM growth-promoting and thrombogenic properties.
Korner
G
G
Department of Radiation and Clinical Oncology, Hadassah University Hospital, Jerusalem, Israel.
Bjornsson
T D
TD
Vlodavsky
I
I
D000042:Absorption; D000818:Animals; D001011:Aorta; D002417:Cattle; D002478:Cells, Cultured; D004728:Endothelium, Corneal / Q000737:chemistry / Q000201:enzymology*; D004730:Endothelium, Vascular / Q000737:chemistry / Q000201:enzymology*; D005109:Extracellular Matrix / Q000737:chemistry / Q000201:enzymology*; D005966:Glucuronidase; D006026:Glycoside Hydrolases / Q000378:metabolism; D008970:Molecular Weight; D017395:Plasminogen Activator Inhibitor 1 / Q000032:analysis; D010959:Tissue Plasminogen Activator / Q000737:chemistry / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000737:chemistry / Q000378:metabolism*; D000099175:Heparanase
D005966:Glucuronidase; D006026:Glycoside Hydrolases; D017395:Plasminogen Activator Inhibitor 1; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; D000099175:Heparanase