Dissociation of alpha 2-plasmin-inhibitor-plasmin complex and regeneration of plasmin activity by SDS treatment.
In order to understand the mechanism for the complex between alpha 2-plasmin-inhibitor (alpha 2-PI) and plasmin to express its specific activity on fibrin autography after SDS-PAGE, we analyzed the effects of SDS on alpha 2-PI molecule and alpha 2-PI-plasmin complex. Treatment of alpha 2-PI by SDS at the concentrations of 0.01% and 0.1% abolished the activity of alpha 2-PI to form a stoichiometric complex with plasmin, whereas it did not interfere with plasmin's activity. More interestingly, in the case of 0.01% SDS, alpha 2-PI was further cleaved to a smaller molecule. Treatment of previously formed alpha 2-PI-plasmin complex by SDS at the concentrations of both 0.01% and 0.1% dissociated the complex and expressed specific amidolytic activity against tripeptide substrate (S-2251), which activity was totally quenched by aprotinin. When alpha 2-PI-plasmin complex was treated by higher concentration of SDS for 12 hours, dissociated free plasmin's band could be observed on SDS-PAGE analysis. It is likely, therefore, that the exposure of alpha 2-PI-plasmin complex to SDS during the procedure of SDS-PAGE dissociates the complex and expresses its specific proteolytic activity in fibrin autography. These features of alpha 2-PI and its complex with plasmin are similar to those of plasminogen activator inhibitor type 1 (PAI-1) and its complex with plasminogen activators (PAs), thus they may represent some common features of the SERPINS.
Department of Physiology, Hamamatsu University, School of Medicine, Shizuoka-ken, Japan.
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Yan
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Urano
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Takada
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Takada
D000933:Antifibrinolytic Agents / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005341:Fibrinolysin / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D009097:Multienzyme Complexes / Q000737:chemistry; D015843:Serpins / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D000979:alpha-2-Antiplasmin
D000933:Antifibrinolytic Agents; D009097:Multienzyme Complexes; D015843:Serpins; D000979:alpha-2-Antiplasmin; C037742:plasmin-plasmin inhibitor complex; D012967:Sodium Dodecyl Sulfate; D005341:Fibrinolysin