title: Plasminogen-activator in human early milk: its partial purification and characterization.
abstract: Milk plasminogen-activator was partially purified from human transitional milk collected at about 10 days after delivery, by a five-step procedure involving chloroform treatment, ammonium sulfate precipitation, and column chromatography on Sephadex G-150, CM Sephadex C-50 and DEAE Sephadex A-50. This gave milk-activator with a maximum purification factor of about 2,400-fold with respect to the skimmed milk. The CM Sephadex-step preparation showed, on polyacrylamide gel electrophoresis, a single plasminogen-activator activity band located between the bands of albumin and prealbumin of human serum. This preparation exhibited no kinin forming activity. The activator hydrolyzed acetyl-glycyl-L-lysine methyl ester with similar order kinetic constants to urokinase, and was inhibited strongly by diisopropylfluorophosphate. The molecular weight of the activator as estimated by gel filtration was approximately 86,000, the isoelectric points as estimated by gel isoelectric focusing were pH 7.2, 6.9 and 6.6, and the activator activity was not quenched by antiurokinase globulin, indicating that the milk-activator is a different entity from urokinase.
mesh_terms: D000645:Ammonium Sulfate / Q000494:pharmacology; D001120:Arginine / Q000031:analogs & derivatives / Q000378:metabolism; D002725:Chloroform / Q000494:pharmacology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007525:Isoelectric Focusing; D007531:Isoflurophate / Q000494:pharmacology; D007705:Kinins / Q000096:biosynthesis; D008895:Milk, Human; D008970:Molecular Weight; D010960:Plasminogen Activators / Q000302:isolation & purification*; D014568:Urokinase-Type Plasminogen Activator / Q000276:immunology
chemical_list: D007705:Kinins; D007531:Isoflurophate; C026512:arginine methyl ester; D002725:Chloroform; D001120:Arginine; D010960:Plasminogen Activators; D014568:Urokinase-Type Plasminogen Activator; D000645:Ammonium Sulfate