Rodent kinin-forming enzyme systems—I

摘要: Low molecular weight (LMW) kininogen was purified 70-fold with a 16% yield from fresh rat plasma by DEAE-Sephadex chromatography, ammonium sulfate precipitation, Sephadex G-200 gel filtration, SP-Sephadex CM-cellulose and filtration. Ferguson plots of polyacrylamide electrophoretic patterns revealed four bands relative weights 64,000, 123,500, 252,436 357,900 (ratio 1:2:4:6). Sodium dodecyl (SDS)-polyacrylamide electrophoresis provided single protein band 72,000, suggesting that the had been caused aggregation oligomeric protein. The LMW Fraction B (3.9 micrograms bradykinin/mg) used to elicit an antiserum in rabbit. Monospecificity demonstrated immunoelectrophoresis (Laurell rocket Grabar methods) and, thus, homogeneity also. (both Fractions A B) formed kinin human urinary kallikrein, kallikrein hog pancreatic kallikrein. Murphy-Sturm lymphosarcoma acid protease also when incubated at pH 3.0. isoelectric point for both fractions 4.3. Amino analyses showed two be rich acidic amino acids have total carbohydrate content 8.5% consisting galactose (1.2 1.5%), mannose (1.9 2.1%), N-acetylglucosamine (4.3 5.1%), N-acetylgalactosamine (0.3%), sialic (0.68%).