The active and the inactive plasminogen activator inhibitor from human endothelial cell conditioned medium are immunologically and functionally related to each other.
In human endothelial cell conditioned medium a fast-acting inhibitor of tissue-type plasminogen activator and urokinase has been detected. Moreover, an inactive inhibitor of these plasminogen activators is present, that can be activated by denaturing agents such as sodium dodecyl sulphate (SDS). The mutual relationship between these inhibitors was studied. The fast-acting plasminogen activator inhibitor from human endothelial cell conditioned medium was purified in a complex with tissue-type plasminogen activator by immune adsorption, using an immobilized anti-tissue-type plasminogen activator antibody. With the complex as an antigen, specific antibodies were raised against this inhibitor in rabbits. The antiserum immunoreacted with both the inactive and the fast-acting plasminogen activator inhibitor. Endothelial cell conditioned medium (containing the inactive plasminogen activator inhibitor) was treated with SDS and the inhibitory activity that emerged was purified. The SDS-generated product formed complexes with tissue-type plasminogen activator with the same molecular mass as those formed with the fast-acting inhibitor. Moreover, the inhibitory activity generated by SDS treatment showed the same kinetic behaviour with tissue-type plasminogen activator as did the fast-acting inhibitor. These data show that the fast-acting and the inactive plasminogen activator inhibitor are immunologically and functionally related to each other, and probably represent different molecular forms of the same protein.
D000918:Antibody Specificity; D002478:Cells, Cultured; D003429:Cross Reactions; D003470:Culture Media / Q000032:analysis; D004727:Endothelium / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis*; D006801:Humans; D005779:Immunodiffusion; D007525:Isoelectric Focusing; D008970:Molecular Weight; D015849:Plasminogen Inactivators; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors
D003470:Culture Media; D006023:Glycoproteins; D015849:Plasminogen Inactivators; D012967:Sodium Dodecyl Sulfate; D010959:Tissue Plasminogen Activator; D014568:Urokinase-Type Plasminogen Activator