Platelet-bound prekallikrein promotes pro-urokinase-induced clot lysis: a mechanism for targeting the factor XII dependent intrinsic pathway of fibrinolysis.
Clots formed from platelet rich plasma were found to be lysed more readily by low concentrations of pro-urokinase (pro-UK) than clots formed from platelet poor plasma. This was not a non-specific effect since the reverse occurred with tissue plasminogen activator. A mechanical explanation due to platelet-mediated clot retraction was excluded by experiments in which retraction was inhibited with cytochalasin B. Therefore, a platelet-mediated enzymatic mechanism was postulated to explain the promotion of fibrinolysis. Casein autography of isolated platelets revealed a approximately 90 kDa band of activity which comigrated with plasma prekallikrein (PK)/kallikrein, a known activator of pro-UK. Furthermore, treatment of platelets with plasma PK activator (PPA), consisting essentially of factor XIIa, induced activation of pro-UK and of chromogenic substrate for kallikrein (S-2302). This activity corresponded to approximately 40-200 pM kallikrein per 10(8) washed and gel filtered platelets per ml. The activation of pro-UK by PPA-pretreated platelets was dose-dependent and inhibited by soybean trypsin inhibitor but not by bdellin, a specific inhibitor of plasmin, nor by the corn inhibitor of factor XIIa. Kinetic analysis of pro-UK activation by kallikrein showed promotion of the reaction by platelets. The KM of the reaction was reduced by platelets by approximately 7-fold, while the kcat was essentially unchanged. In conclusion, PK was shown to be tightly associated with platelets where it can be activated by factor XIIa during clotting. The activation of pro-UK by platelet-bound kallikrein provides an explanation for the observed platelet mediated promotion of pro-UK-induced clot lysis.(ABSTRACT TRUNCATED AT 250 WORDS)
Loza
J P
JP
Department of Medicine, Deaconness Hospital, Harvard Medical School, Boston, MA 02215.
Gurewich
V
V
Johnstone
M
M
Pannell
R
R
D000328:Adult; D001792:Blood Platelets / Q000737:chemistry / Q000502:physiology*; D002863:Chromogenic Compounds; D003018:Clot Retraction; D004789:Enzyme Activation / Q000187:drug effects; D015956:Factor XIIa / Q000502:physiology*; D005342:Fibrinolysis / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D010958:Plasminogen / Q000494:pharmacology; D011288:Prekallikrein / Q000032:analysis / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000494:pharmacology; D013268:Stimulation, Chemical; D010959:Tissue Plasminogen Activator / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000494:pharmacology*
D002863:Chromogenic Compounds; D011994:Recombinant Proteins; D010958:Plasminogen; D011288:Prekallikrein; D015956:Factor XIIa; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator; C067551:saruplase