The isolation and characterization of a ternary human plasmin B-chain-streptokinase-plasminogen complex.
A ternary equimolar human plasmin B-chain-streptokinase-plasminogen complex was isolated from a mixture of the plasmin B-chain-streptokinase complex and human plasminogen at 0 degrees C and 37 degrees C. A ternary complex which was shown to be species specific, was identified and characterized by ultracentrifugal, acrylamide gel electrophoretic, and agarose double diffusion analyses. When mixed at a 1:1 molar ratio at 0 degrees C, 39.9% of the preparation existed as a plasmin B-chain-streptokinase-plasminogen complex; when mixed at 37 degrees C, 86.4% existed as a complex, which was identified by electrophoretic analyses to be a plasmin B-chain-streptokinase-plasmin complex. Sedimentation velocity analyses gave s degrees 20,w values of 3.79 for the plasmin B-chain-streptokinase complex, 4.10 for Lys-plasmin, and 6.23 for the plasmin B-chain-streptokinase-plasmin complex. Sedimentation equilibrium analyses gave molecular weights of 73,900 for the plasmin B-chain-streptokinase complex, 82,900 for Lys-plasmin, and 153,100 for the plasmin B-chain-streptokinase-plasmin complex. The diisopropylphosphorofluoridate (DFP)-inhibited and the p-nitrophenyl-p-guanidino-benzoate (NPGB)-inhibited plasmin B-chain-streptokinase complexes both retained their ability to form a ternary complex with human plasminogen, but this complex did not convert to a plasmin B-chain-streptokinase-plasmin complex. Thus, the active site serine residue is essential for the activator activity of the plasmin B-chain-streptokinase complex, but it is not necessary for the binding of the plasmin B-chain-streptokinase complex to plasminogen to form a ternary complex.
D001665:Binding Sites; D004591:Electrophoresis, Polyacrylamide Gel; D005341:Fibrinolysin / Q000302:isolation & purification*; D006801:Humans; D005779:Immunodiffusion; D008968:Molecular Conformation; D010958:Plasminogen / Q000302:isolation & purification*; D013300:Streptokinase / Q000302:isolation & purification*
D010958:Plasminogen; D013300:Streptokinase; D005341:Fibrinolysin