Differential reactivity of Glu-Gly-Arg-CH2Cl, a synthetic urokinase inhibitor, with single-chain and two-chain forms of urokinase-type plasminogen activator.
Glu-Gly-Arg-CH2Cl, a synthetic inhibitor which alkylates the active-site histidine of urokinase with an apparent second-order rate constant of approximately 10(4) M-1 s-1, reacts differently with single-chain and two-chain forms of urokinase-type plasminogen activators (scu-PA and tcu-PA). Kinetic analysis indicated that the plasminogen activating potential of tcu-PA is irreversibly inhibited in a two-step reaction according to (formula; see text) with Ki = 5.0 X 10(-6) M and k2 = 0.05 s-1. The plasminogen-activating potential of scu-PA, however, is competitively inhibited by Glu-Gly-Arg-CH2Cl with Ki = 1.3 X 10(-6) M. Reversibility of the interaction of Glu-Gly-Arg-CH2Cl with scu-PA was confirmed by the restoration of full enzymatic activity after removal of inhibitor. The differential interaction of Glu-Gly-Arg-CH2Cl with scu-PA and tcu-PA supports the hypothesis that these molecular forms of urokinase-type PA have intrinsically different enzymatic properties.
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology*; D001665:Binding Sites; D005342:Fibrinolysis / Q000187:drug effects; D006639:Histidine; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors*
D000590:Amino Acid Chloromethyl Ketones; D006639:Histidine; C051330:glutamyl-glycyl-arginine chloromethyl ketone; D014568:Urokinase-Type Plasminogen Activator