Inactivation of plasminogen activator inhibitor by oxidants.
The rapidly acting plasminogen activator inhibitor (PAI) purified from cultured bovine aortic endothelial cells (BAEs) was inactivated during iodination with chloramine T and other oxidizing iodination systems. Inactivation was observed in the absence of iodine, suggesting that the loss of activity resulted from the oxidizing conditions employed. In an attempt to further study the nature of this inactivation, the PAI was treated with chloramine T under conditions that specifically oxidize methionine and cysteine residues. Both PAI inhibitory activity and the ability of the PAI to form complexes with tissue-type PA were decreased in a dose-dependent manner by such treatment. The PAI was more sensitive to oxidative inactivation than urokinase, elastase, and alpha 1-protease inhibitor. Incubation of the chloramine T inactivated PAI with methionine sulfoxide peptide reductase in the presence of dithiothreitol (DTT) restored more than 90% of the PAI activity. The reductase is a DTT-dependent enzyme that specifically converts methionine sulfoxide to methionine. Little activity was restored by either the reductase or DTT alone. These results indicate that the oxidation of at least one critical methionine residue is responsible for the loss of PAI activity upon iodination. In this respect, the BAE PAI resembles alpha 1-protease inhibitor, a well-characterized elastase inhibitor that also is inactivated by oxidants. Both inhibitors are members of the serine protease inhibitor superfamily (Serpins), and both have a methionine residue in their reactive center.
D000818:Animals; D001011:Aorta / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D002700:Chloramines / Q000494:pharmacology*; D004727:Endothelium / Q000378:metabolism; D006023:Glycoproteins / Q000037:antagonists & inhibitors* / Q000302:isolation & purification; D007700:Kinetics; D010084:Oxidation-Reduction; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors*; D015849:Plasminogen Inactivators; D014105:Tosyl Compounds
D002700:Chloramines; D006023:Glycoproteins; D015849:Plasminogen Inactivators; D014105:Tosyl Compounds; C016300:chloramine-T; D010960:Plasminogen Activators