Plasminogen activator inhibitor type-1 : reactive center and amino-terminal heterogeneity determined by protein and cDNA sequencing

• Published in: FEBS letters Vol. 209; no. 2; pp. 213 - 218
• Main Authors: Andreasen, P.A., Riccio, A., Welinder, K.G., Douglas, R., Sartorio, R., Nielsen, L.S., Oppenheimer, C., Blasi, F., Danø, K.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15-12-1986; Elsevier
• Subjects: Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Base Sequence; Binding Sites; Biological and medical sciences; cDNA; Denhardt's solution, 0.02 Ficoll, 0.02 polyvinylpyrrolidone, 0.02 bovine serum albumin; DNA - analysis; DNA Restriction Enzymes; DTT, dithiothreitol; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Humans; Hydrolases; man; Nucleotide sequence; PAGE, polyacrylamide gel electrophoresis; PAI-1, type-1 plasminogen activator inibitor; PAI-2, type-2 plasminogen activator inhibitor; PBS, 0.01 M sodium phosphate, pH 7.4, 0.15 M NaCl; Plasminogen - metabolism; Plasminogen activator; Plasminogen activator inhibitor; Protease Inhibitors - pharmacology; PTH, phenylthiohydantoin; Serpin; SSC, 0.15 M NaCl, 0.015 M sodium citrate, pH 7.6; Substrate Specificity; t-PA, tissue-type plasminogen activator; Tissue Plasminogen Activator - genetics; u-PA, urokinase-type plasminogen activator; Urokinase-Type Plasminogen Activator - genetics; Nucleotide sequence; PBS, 0.01 M sodium phosphate, pH 7.4, 0.15 M NaCl; SSC, 0.15 M NaCl, 0.015 M sodium citrate, pH 7.6; Amino acid sequence; Plasminogen activator; t-PA, tissue-type plasminogen activator; PAI-1, type-1 plasminogen activator inibitor; DTT, dithiothreitol; Serpin; Denhardt's solution, 0.02 mg ml Ficoll, 0.02 mg ml polyvinylpyrrolidone, 0.02 lmg ml bovine serum albumin; u-PA, urokinase-type plasminogen activator; PAGE, polyacrylamide gel electrophoresis; PTH, phenylthiohydantoin; Plasminogen activator inhibitor; PAI-2, type-2 plasminogen activator inhibitor; Human; Protein inhibitor; Complementary DNA; Gene; Enzyme; N terminal-Sequence
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(86)81113-9

Both the urokinase-type and tissue-type plasminogen activator can convert their 5̃4 kDa type-1 inhibitor (PAI-1) to an inactive form with a lower apparent molecular mass. We have determined the amino-terminal amino acid sequences of human native and converted PAI-1, and isolated PAI-1 cDNA and determined the nucleotide sequence in regions corresponding to the amino-terminus and the cleavage site. The data show that the conversion of the inhibitor consists of cleavage of an Arg-Met bond 33 residues from the carboxy-terminus, thus localizing the reactive center of the inhibitor to that position. In addition, a heterogeneity was found at the amino-terminus, with a Ser-Ala-Val-His-His form and a two-residue shorter form (Val-His-His-) occurring in approximately equal quantities.