High-level expression of active human plasminogen activator inhibitor type 1 (PAI-1) in E. coli.

Abstract

Plasminogen activator inhibitor 1 (PAI-1) is an important regulator of plasminogen activation, which inhibits both tissue-type plasminogen activator (tPA) and urokinase-type plasminogen activator (uPA). The DNA sequence encoding mature PAI-1 protein was inserted into an inducible expression vector. This gene was highly expressed to produce a soluble active protein in E. coli cells. The amount of the recombinant protein was up to 20% of total cellular protein. By efficient purification with a yield of about 15-20%, the recombinant protein could be purified to homogeneity with its specific activity up to 6.1 x 10(4) (uPA) IU/mg. Its inhibitory activity declined during incubation at 37 degrees C with a half life of about 2 hr.