Characterization of plasminogen activator produced by an established cell line from human ovary.
An established cell line (OC-1) was obtained from human ovarian tissue, which yielded a high concentration of plasminogen activator (PA) in the culture medium. The PA (OC-1-PA) produced by the cell line was purified and compared with urokinase (UK), proform of UK (pro-UK), and tissue-type PA (t-PA) purified from human melanoma cells (Bowes). OC-1-PA was purified by Zn chelate-Sepharose affinity chromatography followed by high-performance liquid chromatography with a Zn chelate-5PW column and with a p-amino-benzamidine-5PW column, giving a yield of 58.3% and a purification factor of 15,439. This purified material revealed a single band of Mr 55,000 on sodium dodecylsulfate polyacrylamide gel electrophoresis in the presence or absence of reducing agents. Electrophoretic enzymography demonstrated that the Mr 55,000 protein band had a plasminogen-dependent fibrinolytic activity. Treatment with plasmin did not change the Mr even in the presence of reducing agents. These results suggest that OC-1-PA has a single-chain structure protected from protease degradation, which is completely different from UK. The activator had higher affinities for lysine and fibrin than those of UK or pro-UK. An immunological study demonstrated that OC-1-PA cross-reacted with anti-UK IgG but not with anti-t-PA IgG. All these findings indicate that OC-1-PA belongs immunologically to the UK type, but its structure differs from that of UK.
D002460:Cell Line; D055598:Chemical Phenomena; D002621:Chemistry; D005260:Female; D005337:Fibrin / Q000378:metabolism; D006801:Humans; D007526:Isoelectric Point; D008239:Lysine / Q000378:metabolism; D010053:Ovary / Q000166:cytology / Q000378:metabolism*; D010960:Plasminogen Activators / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism
D005337:Fibrin; D010960:Plasminogen Activators; D008239:Lysine