The cleavage and inactivation of plasminogen activator inhibitor type 1 and alpha2-antiplasmin by reptilase, a thrombin-like venom enzyme.
Reptilase, defibrase and ancrod are thrombin-like venom enzymes that cleave fibrinogen to release fibrinopeptide-A and generate fibrin monomers. Although these enzymes decrease fibrinogen levels in vivo, presumably by enhancing fibrinolytic activity, the mechanism has not been identified. In the present study, we analyzed their effects on the inhibitors of fibrinolysis. Plasminogen activator inhibitor-1 (PAI-1) was cleaved at its C-terminus by reptilase and lost its specific activity. Alpha2-antiplasmin (alpha2-AP) was cleaved both at the Pro19-Leu20 peptide bond and at its C-terminus by reptilase, and also lost its specific activity. The apparent second-order rate constants (mol/l per min per Batroxobin unit) were 0.22 for the cleavage of PAI-1 (3.2 micromol/l) and 0.19 for that of alpha2AP (6.4 micromol/l), which were approximately 200-fold lower than that (47.0) for the cleavage of fibrinogen (1.1 micromol/l). Neither defibrase nor ancrod cleaved and inactivated these inhibitors. Only reptilase enhanced euglobulin clot lysis in vitro at high concentration, due probably to PAI-1 inactivation. Since all these three enzymes enhance fibrinolysis similarly during defibrination therapy, the neutralization or inactivation of the inhibitors of fibrinolysis appeared not to represent the main mechanism for the enhancement.
Urano
T
T
Department of Physiology, Hamamatsu University, School of Medicine, Shizuoka, Japan.
Ihara
H
H
Takada
Y
Y
Fujie
M
M
Takada
A
A
D000721:Ancrod / Q000494:pharmacology; D012103:Batroxobin / Q000378:metabolism* / Q000494:pharmacology; D001777:Blood Coagulation / Q000187:drug effects; D010450:Endopeptidases / Q000494:pharmacology; D005342:Fibrinolysis / Q000187:drug effects; D005343:Fibrinolytic Agents / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D010446:Peptide Fragments; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism*; D012712:Serum Globulins / Q000494:pharmacology; D013917:Thrombin; D014688:Venoms; D000979:alpha-2-Antiplasmin / Q000187:drug effects / Q000378:metabolism*
D005343:Fibrinolytic Agents; D010446:Peptide Fragments; D017395:Plasminogen Activator Inhibitor 1; D012712:Serum Globulins; D014688:Venoms; D000979:alpha-2-Antiplasmin; D010450:Endopeptidases; D000721:Ancrod; D012103:Batroxobin; D013917:Thrombin