Platelet plasminogen activator inhibitor: purification and characterization of interaction with plasminogen activators and activated protein C.
Plasminogen activator inhibitor (PAI) was purified in active form from porcine platelets under nondenaturing conditions. The purified inhibitor (Mr 47,000) reacts with tissue-type plasminogen activator (t-PA), urokinase (UK), and activated protein C (APC) to yield both SDS-stable complexes and a modified PAI of slightly reduced molecular weight. The second-order rate constants for the inhibition of t-PA and UK by PAI are 3.5 X 10(7) and 3.4 X 10(7) M-1 s-1, respectively. Activated protein C reacts with PAI with a second-order rate constant of 1.1 X 10(4) M-1 s-1. This rate is not accelerated by protein S, phospholipid, and calcium, or heparin. It is concluded that (1) PAI can function as both inhibitor and substrate of its target proteases, (2) if APC promotes fibrinolysis via inactivation of PAI, then APC must be present in concentrations several orders of magnitude greater than t-PA, or the interaction of APC and PAI must be accelerated by presently unknown mechanisms, and (3) in the absence of heparin, platelet PAI is the most rapid inhibitor of APC yet described.
Fay
W P
WP
Department of Internal Medicine, Mayo Clinic and Foundation, Rochester, Minnesota 55905.
Owen
W G
WG
D000818:Animals; D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism*; D006023:Glycoproteins / Q000097:blood*; D066298:In Vitro Techniques; D007700:Kinetics; D008970:Molecular Weight; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors* / Q000378:metabolism; D015849:Plasminogen Inactivators; D011486:Protein C / Q000037:antagonists & inhibitors / Q000378:metabolism; D013552:Swine
D006023:Glycoproteins; D015849:Plasminogen Inactivators; D011486:Protein C; D010960:Plasminogen Activators