[The interaction of human alpha 1-antitrypsin with human plasmin].
The interaction of alpha 1-antitrypsin (alpha 1-AT) with plasmin was investigated, and the molecular weight of the inhibitor was also re-evaluated. The value of molecular weight of alpha 1-AT determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) method showed a difference depending on the presence or absence of the reducing agent, resulting in 72,000 dalton before reduction and 59,000 dalton after reduction. Conclusively, the molecular weight of alpha 1-AT was appropriate to be 59,000 dalton from considering the molecular shape of the protein. The interaction of alpha 1-AT with plasmin was analysed by SDS-PAGE method. Unreduced analysis revealed that two kinds of complexes with different molecular weight (the major of 155,000 dalton and the minor of 140,000 dalton) were formed time dependently, suggesting that the former was a native complex and the latter was a degraded product. Reduced analysis disclosed that the light chain of plasmin involved the complex formation with the inhibitor, and a peptide of 16,000 dalton appeared during the reaction. From these observations, the mechanism of action was summarized as follows. First, alpha 1-AT inhibited all of the plasmin activities by forming a 1: 1 stoichiometric complex with the enzyme, presumably with the active center of the enzyme, whose complex is undissociable in the presence of denaturing or reducing agents or both. Secondly, the native complex broke into a degraded product and a released peptide by limited proteolysis with the free plasmin which existed in the reaction mixture even with an excess of alpha 1-AT due to the reaction of complex formation being time consuming. The clinical significance of alpha 1-AT on fibrinolysis was also subject for discussion.
D004591:Electrophoresis, Polyacrylamide Gel; D005341:Fibrinolysin / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D006801:Humans; D008970:Molecular Weight; D000515:alpha 1-Antitrypsin / Q000032:analysis / Q000302:isolation & purification / Q000494:pharmacology*
D000515:alpha 1-Antitrypsin; D005341:Fibrinolysin