A chromogenic enzymatic assay capable of detecting prourokinase-like material in plasma

P. R. Kebabian*, J. Henkin

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    5 Scopus citations

    Abstract

    We report a functional assay capable of quantifying prourokinase (ProUK)- like material in plasma where urokinase (UK) is also present. The assay involves inactivation of urokinase with a specific, active site directed irreversible inhibitor, dansyl-glutamyl glycyl arginine chloromethylketone (dansyl- GGACK). Excess inhibitor is subsequently quenched with dithiothreitol (DTT). The ProUK-like material in plasma is then converted to active urokinase with thermolysin, a proteolytic enzyme of bacterial origin. Alpha 2-macroglobulin in plasma inhibits thermolysin; however alpha 2-macroglobulin is inactivated with methylamine. The assay can detect as little as 20 ng of ProUK and is linear from 20 to 120 ng. The assay was applied to quantify the amount of ProUK-like material in plasma obtained from dog at various times after i.v. administration of 100,000 or 75,000 U/kg, of pro-urokinase.

    Original languageEnglish (US)
    Pages (from-to)401-407
    Number of pages7
    JournalThrombosis Research
    Volume65
    Issue number3
    DOIs
    StatePublished - Feb 1 1992

    Keywords

    • Prourokinase
    • Urokinase
    • alpha-2 macroglobulin
    • thermolysin

    ASJC Scopus subject areas

    • Hematology

    Fingerprint

    Dive into the research topics of 'A chromogenic enzymatic assay capable of detecting prourokinase-like material in plasma'. Together they form a unique fingerprint.

    Cite this