Activation of single-chain urokinase-type plasminogen activator by a hemorrhagic metalloproteinase, jararafibrase I, in Bothrops jararaca venom.
Urokinase-type plasminogen activator (uPA) activates plasminogen to plasmin, which is involved in the degradation of the vascular basement membrane and extracellular matrix. The present study was undertaken to examine the effects of several hemorrhagic metalloproteinases, jararafibrase (JF) I, II, III and IV, purified from Bothrops jararaca venom, on the single-chain zymogen form of uPA (scuPA). Activation of scuPA by JF I IV was estimated using a synthetic substrate for uPA (S-2444). Only JF I activated the scuPA in a time- and dose-dependent manner. SDS-PAGE analysis revealed that, after incubation with JF I, the intensity of the 55 kDa band of scuPA decreased concomitantly with increases in the intensity of the major two bands at 32 and 22 kDa under reduced and non-reduced conditions. The 32 kDa band demonstrated fibrinolytic activity in fibrin-zymographic studies. Amino-acid-sequence analysis revealed that JF I cleaved the position of 143Glu-144Leu in scuPA, indicating that JF I formed low molecular weight scuPA. From these results, it seems possible that activation of scuPA by JF I could be responsible in part for the local hemorrhage and tissue damage that are frequently observed in human victims of B. jararaca envenoming.
Department of Physiology, Miyazaki Medical College, Kiyotake, Japan.
M
M
Sugiki
E
E
Yoshida
K
K
Anai
M
M
Maruyama
D003435:Crotalid Venoms / Q000737:chemistry / Q000494:pharmacology*; D004789:Enzyme Activation; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D008666:Metalloendopeptidases / Q000302:isolation & purification / Q000494:pharmacology*; D010960:Plasminogen Activators / Q000737:chemistry / Q000378:metabolism*
D003435:Crotalid Venoms; D005343:Fibrinolytic Agents; D010960:Plasminogen Activators; D008666:Metalloendopeptidases; C076581:jararafibrase I