Kinetic analysis of plasminogen activation by staphylokinase/plasminogen complex in the presence of fibrin.
Staphylokinase (SAK) expresses plasminogen activator activity by forming a complex with plasminogen. In order to elucidate the mechanism for the expression of enzymatic activity of the complex, a cross-linked staphylokinase/plasminogen (SAK/plg) complex was produced with disuccinimidyl suberate, and its enzymatic characteristics were compared with those of a streptokinase/plasminogen (SK/plg) complex. SAK/plg complex and SK/plg complex showed a band with a molecular weight of 110 kDa and 140 kDa by SDS-PAGE under non-reduced condition, respectively. Both complexes exhibited plasminogen activator activity in a concentration-dependent manner on fibrin film and synthetic chromogenic substrate assay. The kinetic analysis of enzymatic activity of both complexes was performed. The plasminogen activator activity of SAK/plg complex was enhanced about 5-fold in the presence of FCB-2. However, SK/plg complex showed only 1.7-fold increase in the presence of FCB-2. These findings indicate that the SAK/plg complex reacts with fibrin, and efficient plasminogen activation is induced on fibrin surface.
Okada
K
K
Department of Physiology, Kinki University School of Medicine, Osakasayama, Japan.
Yuasa
H
H
Hagiya
Y
Y
Fukao
H
H
Ueshima
S
S
Matsuo
O
O
D000595:Amino Acid Sequence; D004789:Enzyme Activation / Q000187:drug effects; D005337:Fibrin / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008666:Metalloendopeptidases / Q000378:metabolism*; D008969:Molecular Sequence Data; D010958:Plasminogen / Q000378:metabolism*
D005337:Fibrin; D010958:Plasminogen; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus