Released protease-nexin regulates cellular binding, internalization, and degradation of serine proteases.
Protease-nexin (PN), a component released by normal human fibroblasts into culture medium, forms covalent linkages with thrombin (Th) and the urinary plasminogen activator urokinase, apparently with their catalytic site serines. The present studies explored the function of PN by examining the interaction of protease-PN complexes with human fibroblasts and the consequences of this interaction. Th-PN and urokinase-PN complexes bind to cells via the PN portion of the complexes. The binding is selectively inhibited by heparin. Because PN has a heparin-binding site, this indicates that protease-PN complexes might bind to a cellular heparin-like site. After binding, the complexes are internalized. By inhibiting endocytosis with phenylarsine oxide, which does not affect cellular binding of Th-PN complexes, we showed that complexes must be internalized before they are degraded. Kinetic analysis of internalization and degradation of Th-PN showed that complexes are internalized more rapidly than they dissociate from the cell surface; by 120 min of incubation at 37 degrees C most cell-bound Th-PN complexes are degraded to amino acids. The results are summarized in a model showing how PN mediates the cellular binding, internalization, and degradation of serine proteases through formation of protease-PN complexes. This series of events may be involved in the regulation of serine protease activity at the cell surface and in the extracellular environment.
D016564:Amyloid beta-Protein Precursor; D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000494:pharmacology*; D002478:Cells, Cultured; D004705:Endocytosis; D010450:Endopeptidases / Q000378:metabolism*; D006493:Heparin / Q000494:pharmacology; D006629:Hirudins / Q000494:pharmacology; D006801:Humans; D008297:Male; D051379:Mice; D011480:Protease Inhibitors / Q000378:metabolism*; D058255:Protease Nexins; D011506:Proteins / Q000378:metabolism*; D011956:Receptors, Cell Surface; D012697:Serine Endopeptidases; D015842:Serine Proteinase Inhibitors; D013917:Thrombin / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
D016564:Amyloid beta-Protein Precursor; D002352:Carrier Proteins; D006629:Hirudins; D011480:Protease Inhibitors; D058255:Protease Nexins; D011506:Proteins; D011956:Receptors, Cell Surface; D015842:Serine Proteinase Inhibitors; D006493:Heparin; D010450:Endopeptidases; D012697:Serine Endopeptidases; D013917:Thrombin; D014568:Urokinase-Type Plasminogen Activator