Sensitive radiochemical esterolytic assays for urokinase.
Two radiochemical esterolytic assays for urokinase are described. One assay is based on the urokinase-dependent hydrolysis of Nalpha-acetyl-glycyl-L-lysine [3H]methyl ester and the other on the urokinase-dependent activation of plasminogen and assay of generated plasmin with Nalpha-tosyl-L-arginine [3H]methyl ester. The assays are performed in tubes placed in liquid scintillation counting vials. At the end of the experiment generated [3H]methanol is extracted into the liquid scintillation cocktail and counted. Unhydrolyzed substrate largely remains in the aqueous phase and contributes only a small fraction of the counts. This facile separation of 3H-labeled alcohol from the ester substrate allows the simple and highly sensitive assay for urokinase. The assays give results in good agreement with the classical fibrin plate assay.
D000328:Adult; D004151:Dipeptides / Q000378:metabolism; D010450:Endopeptidases / Q000032:analysis*; D005260:Female; D005337:Fibrin / Q000378:metabolism; D005341:Fibrinolysin / Q000378:metabolism; D005998:Glycine / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008239:Lysine / Q000378:metabolism; D008297:Male; D000432:Methanol / Q000032:analysis; D013696:Temperature; D014106:Tosylarginine Methyl Ester / Q000378:metabolism; D014316:Tritium; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis* / Q000652:urine
D004151:Dipeptides; D014316:Tritium; D005337:Fibrin; D014106:Tosylarginine Methyl Ester; D010450:Endopeptidases; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; D008239:Lysine; D005998:Glycine; D000432:Methanol