The reciprocal effects of epsilon-aminohexanoic acid and chloride ion on the activation of human [Glu1]plasminogen by human urokinase.

The activation of human [Glu1]plasminogen [( Glu1]Pg) by high-molecular-weight two-chain human urinary urokinase [EC 3.4.21.31) and low-molecular-weight two-chain human urinary urokinase is inhibited by Cl- at physiological concentrations and stimulated by epsilon-aminohexanoic acid (epsilon Ahx; epsilon-aminocaproic acid). The inhibition by Cl- does not occur in the presence of concentrations of epsilon Ahx that saturate the acid's weak binding sites on [Glu1]Pg, and the stimulation by epsilon Ahx is maximally exhibited in the presence of Cl-. We have used intrinsic fluorescence measurements with [Glu1]Pg to show that the conformational alteration and the concomitant increase in activation rate that accompanies epsilon Ahx-binding to [Glu1]Pg in the presence of Cl- does not occur in the same manner without Cl-. Further, the decrease in the intrinsic fluorescence that is attendant to Cl- binding to [Glu1]Pg in the absence of epsilon Ahx is not observed in the presence of this effector molecule. Analyses of the results of this manuscript strongly indicate that a conformation of [Glu1]Pg that is not optimal for its activation by urokinase is adopted in the presence of Cl-, and this is relieved by epsilon Ahx. This has important implications in the inhibition of [Glu1]Pg activation in the solution phase of blood plasma and in the large acceleration of this process when plasminogen is bound to physiological positive effectors via its epsilon Ahx-binding site(s).