Human Plasminogen activator inhibitor-1

Plasminogen activator inhibitor-1 (PAI-1) is a member of the serine protease inhibitor (serpin) superfamily and a central regulatory protein in the blood coagulation system. PAI-1 is unique among serpins in exhibiting distinct active and inactive (latent) conformations in vivo. Human PAI-1 is a single-chain glycoprotein with a molecular weight of 43 kDa. This inhibitor acts as "bait" for tissue-type and urokinase-type plasminogen activators (tPA and uPA) and protein C. Its rapid interaction with tPA may function as a major control point in the regulation of fibrinolysis. The highly mobile reactive-center loop (RCL) is thought to account for both the rapid inhibiton of plasminogen activators, and the rapid and spontaneous transition of the unstable, active form of PAI-1 into the stable, inactive conformation (t1/2 at 37oC, 2 hours). The inactive form can be partially reactivated by denaturants such as urea, guanidine hydrochloride or SDS. High concentrations of PAI-1 have been associated with human thromboembolic disease. PAI-1 activity may limit the extent of tumor metastasis, since uPA activity is a major contributory factor promoting dissolution of tumor matrix and basement membrane.

The sequence corresponding to amino acids 24 to 402 of human PAI-1 (mature form) tagged with 6xHis on the carboxy-terminal was expressed in E. Coli.

To fully activate the latent PAI-1, the PAI-1 must be incubated in Activation Buffer (2X Activation Buffer: 8 M Guanidine HCl, 40 mM Sodium Acetate, pH 5.6, 400 mM NaCl, 0.2% Tween 20) for 15 to 30 minutes (Sancho, 1994).The recombinant PAI-1 is able to bind to uPA resulting in a SDS-stable complex.

Liquid. In 150 mM Na2HPO4, pH 6.6, 500 mM NaCl, 2 mM Glutathione, and 0.01% Tween-80.

Store at -80ºC. Thaw on ice and freeze aliquots at -80ºC for longer storage. This product is stable for at least 2 years as supplied. Avoid repeated freeze/thaw cycles.

Specific Activity: By uPA activity assay, the recombinant PAI-1 contains a mixture of the active and latent forms, with >45% of active form

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