Influence of various structural domains of fibrinogen and fibrin on the potentiation of plasminogen activation by recombinant tissue plasminogen activator.
Fibrinogen, fibrin, and related fragments have varying stimulatory effects on the initial rate of the activation of human plasminogen ([ Glu1]Pg) by recombinant tissue plasminogen activator (rt-PA). A detailed analysis of this enhancement was undertaken using various purified and complexed forms of the known domains of fibrin(ogen) with a view to gaining additional knowledge regarding the substructures of fibrinogen and fibrin that are important for their stimulatory capacities. Both arvin-mediated fibrin, as well as fibrinogen fragments generated as a result of its cleavage with CNBr, stimulate the activation in a biphasic manner, most likely as a result of changes in the promoter molecule accompanying the denaturation processes that are normally employed to either solubilize or generate these particular promoters. Using purified fibrinogen and fibrin fragments, it was found that fragment E, which binds to [Glu1]Pg, does not enhance the activation reaction, while fragment D1 has a potentiating effect. This suggests that the binding of [Glu1]Pg to fibrin(ogen) alone is not, in itself, sufficient for stimulation of activation to occur, but that the rt-PA-fibrin(ogen) interaction is fundamental to this same process. All purified and mixtures of fragments containing the fragment D domain (e.g., D2E, X-oligomer, fragment X) stimulate the reaction to a greater degree than fibrinogen and fragment D1. It is concluded that the fibrinogen D domain is a sine qua non for the enhancement reaction, while structures containing the E domain had a symbiotic effect on enhancement.
de Serrano
V S
VS
Department of Chemistry, University of Notre Dame, Indiana 46556.
Urano
T
T
Gaffney
P J
PJ
Castellino
F J
FJ
D001665:Binding Sites; D002384:Catalysis; D004357:Drug Synergism; D005337:Fibrin / Q000032:analysis*; D005340:Fibrinogen / Q000032:analysis*; D005341:Fibrinolysin / Q000032:analysis; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D010446:Peptide Fragments / Q000494:pharmacology; D010958:Plasminogen / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D013053:Spectrophotometry; D010959:Tissue Plasminogen Activator / Q000235:genetics / Q000494:pharmacology*
D010446:Peptide Fragments; D011994:Recombinant Proteins; D005337:Fibrin; D005340:Fibrinogen; D010958:Plasminogen; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin