Isolation of tissue plasminogen activator from skin lesions with allergic vasculitis

A tissue plasminogen activator was extracted from skin lesions with allergic vasculitis and purified by successive column chromatography on Sephadex G-200, DEAE-cellulose, Hydroxyaptite-cellulose and polyacrylamide gel electrophoresis. By these procedures, 160 micrograms of enzyme with a specific activity of 843.8 international units/mg protein was obtained from 5 g of original skin. The purified material was homogeneous as ascertained by sodium dodecyl sulfate polyacrylamide gel electrophoresis and had an apparent molecular weight of 110,000 as measured by gel filtration on Sephadex G-200. Its identity with human urokinase was investigated and was found to possess the same plasminogen activator activity as that of urokinase. It had high amindolytic activity, but only slight N-alpha-acetyl-glycyl-L-lysine methyl ester esterolytic activity. This tissue plasminogen activator was confirmed to be immunologically identical to human urokinase.