Binding of tissue-type plasminogen activator to human melanoma cells.
We have shown (Bizik et al., Cell Regul 1:895-905, 1990) that tPA can activate plasminogen on the surface of human melanoma cells in the presence of alpha 2-macroglobulin (alpha 2M) secretion. In the present study, we investigated the binding of tPA on the surface of Bowes melanoma cells, selected since they lacked production of PAI-1 and alpha 2M. Elution of tPA from the cell layers indicated that polylysine (5 micrograms/ml) and tranexamic acid (10 mM), an analog of lysine, were the most efficient agents for disrupting the interaction between tPA and cell surface component(s). Using a panel of monoclonal antibodies against individual domains of tPA revealed that an antibody directed to the kringle-2 domain of tPA interfered most significantly with cell-surface plasmin generation. As tPA is a glycoprotein, interactions between the tPA sugar moieties and cell surface were also tested by the use of a series of monosaccharides. N-acetyl-D-glucosamine (100 mM) was the most potent sugar to release tPA from melanoma cells, but the results indicated that the oligosaccharides of tPA play only a supportive role in the binding of tPA to the cell surface. Quantitative comparison of the cell surface localized tPA, which was eluted by tranexamic acid, with the total cellular tPA showed that cell surface bound tPA could represent up to 10%. We conclude that tPA interacts with the melanoma cell surface in a similar manner as has been described for binding of tPA to fibrin and to the putative endothelial cell surface receptor.
Department of Virology, University of Helsinki, Finland.
J
J
Bizik
R W
RW
Stephens
M
M
Grofova
A
A
Vaheri
D000911:Antibodies, Monoclonal; D002241:Carbohydrates / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007477:Ions; D008545:Melanoma / Q000378:metabolism* / Q000473:pathology; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis; D011107:Polylysine / Q000494:pharmacology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D014148:Tranexamic Acid / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D007477:Ions; D017395:Plasminogen Activator Inhibitor 1; D011107:Polylysine; D014148:Tranexamic Acid; D010959:Tissue Plasminogen Activator