Efficacy of plasmin, microplasmin, and streptokinase-plasmin complex for the in vitro degradation of fibronectin and laminin- implications for vitreoretinal surgery.
OBJECTIVE
Plasmin enzyme generates vitreoretinal separation by degradation of laminin and fibronectin in the vitreoretinal interface. It can be activated from plasminogen by urokinase, tissue plasminogen activator, or by formation of a 1:1 complex with streptokinase. The latter is then converted into a streptokinase-plasmin-complex (SK-P), which displays fibrinolytic activity and can generate free plasmin by proteolysis of plasminogen. We compared the efficacy of SK-P, SK-P activated plasmin, urokinase activated plasmin (UK-P), and microplasmin, a truncated form of plasmin, in cleaving laminin and fibronectin.


METHODS
Streptokinase (SK) was added to human plasminogen in molar ratios between 1:100 and 2:1, generating SK-P at ratios > 1:1, and mixtures of SK-P and free plasmin (SK-P/plasmin) at lower ratios. SK-PL, SK-P/plasmin, UK-P, and microplasmin were added to laminin and fibronectin, incubated at 37 degrees C for 30 min-22 hr and processed for SDS-PAGE.


RESULTS
Proteolysis using SK-activated plasminogen increased when the SK/plasminogen ratio was decreased, generating increasing amounts of free plasmin. Microplasmin and urokinase-activated plasmin displayed similar proteolysis of both laminin and fibronectin as SK/plasminogen at ratios of 1:10 or lower.


CONCLUSIONS
The mode of plasminogen activation influences the efficacy of proteolysis for laminin and fibronectin and should be considered when plasmin is used in vitreoretinal surgery.
Hermel
M
M
Department of Ophthalmology, University of Aachen, Aachen, Germany. mhermel@ukaachen.de
Dailey
W
W
Hartzer
M K
MK
D004338:Drug Combinations; D004591:Electrophoresis, Polyacrylamide Gel; D005341:Fibrinolysin / Q000494:pharmacology*; D005342:Fibrinolysis; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D010446:Peptide Fragments / Q000494:pharmacology*; D010958:Plasminogen / Q000494:pharmacology*; D012160:Retina / Q000601:surgery; D013300:Streptokinase / Q000494:pharmacology*; D014821:Vitrectomy
D004338:Drug Combinations; D005353:Fibronectins; D007797:Laminin; D010446:Peptide Fragments; C049423:streptokinase-plasminogen complex; C054561:microplasmin; D010958:Plasminogen; D013300:Streptokinase; D005341:Fibrinolysin