title: High-level synthesis of biologically active human plasminogen activator inhibitor type 1 (PAI-1) in Escherichia coli.
abstract: Segments of a cDNA encoding human plasminogen activator inhibitor type 1 (PAI-1) were subcloned into a highly regulated and inducible Escherichia coli expression system. A plasmid encoding the mature form of human endothelial PAI-1 produced a functional recombinant molecule, as indicated by its ability to inhibit tissue plasminogen activator's enzymatic activity. In contrast to PAI-1 isolated from human fibrosarcoma cells, the biological activity of the recombinant PAI-1 was not dependent on pretreatment with denaturing agents. A construct encoding a polypeptide lacking the first 80 amino acids of PAI-1 also produced elevated levels of the truncated recombinant protein. However, this truncated product was functionally inactive, indicating that an intact N terminus is required for activity.
authors: E.I. DuPont de Nemours and Co., Central Research and Development Department, Wilmington, DE 19880.
authors: W P
authors: WP
authors: Sisk
authors: G L
authors: GL
authors: Davis
authors: D
authors: D
authors: Kingsley
authors: A T
authors: AT
authors: Chiu
authors: T M
authors: TM
authors: Reilly
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D015849:Plasminogen Inactivators / Q000378:metabolism* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology
chemical_list: D004274:DNA, Recombinant; D015849:Plasminogen Inactivators; D011994:Recombinant Proteins