Human microplasmin is a catalytically active fragment of human plasmin. It consists of a 31-residue C-terminal peptide derived from the A chain bound through two disulfide bonds to the intact B chain of plasmin. It has similar amidolytic and proteolytic activities as the native human Lys-plasmin on a molar basis. Human microplasmin can form a complex with streptokinase, in a one to one stoichiometry, like the native human Lys-plasmin. The stoichiometric human microplasmin and streptokinase complex is an efficient activator of bovine plasminogen which can not be activated by streptokinase alone. The formation of human microplasmin.streptokinase complex was also directly demonstrated by a gel filtration column chromatography. Moreover, bovine plasminogen can not be activated by a mixture of bovine or porcine microplasmin and streptokinase. The equimolar complex of human microplasmin.streptokinase, human Lys-plasmin.streptokinase, or streptokinase alone has the same activator activity toward human Lys-plasminogen. The human microplasmin.streptokinase complex, however, has a significantly higher activator activity than human Lys-plasmin.streptokinase complex or streptokinase alone toward human Glu-plasminogen. The direct interaction between streptokinase and light chain domain of human plasmin is demonstrated in the complex formation. The difference in the activator activities of plasmins from various animal sources in complex with streptokinase therefore might be due to the difference in the compositions of light chains of plasmins.