Production, purification and characterization of the plasminogen activator in teratocarcinoma stem cells induced with sodium butyrate.
Suspension cultures of pluripotent teratocarcinoma cells were induced with sodium butyrate to produce plasminogen activator (PA), generally regarded as a marker enzyme of differentiation of the teratocarcinoma. The induction of plasminogen activator was very efficient, resulting in production of sufficient enzyme to allow its purification. The activator was inactivated by rabbit anti-human melanoma plasminogen activator antiserum, indicating that it was a tissue-type activator (t-PA). The enzyme was purified by column chromatograph on phosphocellulose, zinc-chelate agarose, Con-A Sepharose and Sephadex G-150. The preparation at the final step of purification gave a single peak of enzyme activity at pH 7.3 +/- 0.1 on isoelectric focusing, and showed a molecular weight of approximately 77,000 on SDS PAGE.
D000818:Animals; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D002460:Cell Line; D002471:Cell Transformation, Neoplastic / Q000187:drug effects / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D054278:Embryonal Carcinoma Stem Cells; D004790:Enzyme Induction; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D008545:Melanoma / Q000201:enzymology; D051379:Mice; D014411:Neoplastic Stem Cells / Q000201:enzymology*; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000302:isolation & purification; D015849:Plasminogen Inactivators; D013234:Stem Cells / Q000201:enzymology*; D013724:Teratoma / Q000201:enzymology*
D002087:Butyrates; D007106:Immune Sera; D015849:Plasminogen Inactivators; D020148:Butyric Acid; D010960:Plasminogen Activators