title: Kinetic studies on the plasminogen activation by the staphylokinase-plasmin complex.
abstract: A pure complex of staphylokinase and plasmin was prepared by affinity chromatography with lysine-Sepharose, which enabled the simple analysis of the mechanism of plasminogen activation by staphylokinase. We used a truncated staphylokinase (SAK), which lacks the 10 amino acid residues at the NH2 terminal of native staphylokinase. The purity of this complex was confirmed by the native PAGE profile. Image analysis of the SDS-PAGE profile revealed that the molar ratio of plasmin and SAK in the complex was about 1:1. Using this SAK-plasmin complex, the kinetic parameters for the activation of Glu- or Lys-plasminogen were determined. The kinetic constant, kcat/Km, obtained when Lys-plasminogen was used as a substrate was approximately 10 times higher than that obtained when Glu-plasminogen was used. This plasminogen activation property of the SAK-plasmin complex was comparable to that of other plasminogen activators, such as streptokinase, urokinase, and tissue-type plasminogen activator (t-PA). This SAK-plasmin complex will simplify the elucidation of plasminogen activation by SAK. Through kinetic studies, the fibrin specificity and participation of plasminogen activator inhibitor will be clarified.
authors.affiliation: Yakult Central INstitute for Microbiological Research, Tokyo.
authors.forename: H
authors.initials: H
authors.lastname: Shibata
authors.forename: M
authors.initials: M
authors.lastname: Nagaoka
authors.forename: M
authors.initials: M
authors.lastname: Sakai
authors.forename: H
authors.initials: H
authors.lastname: Sawada
authors.forename: T
authors.initials: T
authors.lastname: Watanabe
authors.forename: T
authors.initials: T
authors.lastname: Yokokura
mesh_terms: D005341:Fibrinolysin / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D008666:Metalloendopeptidases / Q000737:chemistry*; D015233:Models, Statistical; D010960:Plasminogen Activators / Q000737:chemistry*
chemical_list: D010960:Plasminogen Activators; D005341:Fibrinolysin; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus