The effect of lanthanide ions on enteropeptidase-catalyzed activation of trypsinogen

Rare earths were found to be powerful inhibitors of enteropeptidase-catalyzed (enterokinase, EC 3.4.21.9) activation of trypsinogen. Inhibition was complete at a La3+ concentration of 12.5-10(-6) M in the assay system used and still detectable at a concentration of 1.25-10(-6) M. Inhibition was observed with all lanthanides tested. No significant differences between individual metals could be established under the conditions of the inhibition assay. Increasing ionic strength decreased enzyme activity and progressively diminished the inhibitory effect of rare earth ions suggesting an electrostatic basis for the mechanism of this inhibition. La3+ did not significantly affect enteropeptidase-mediated hydrolysis of N-benzoyl-L-arginine ethyl ester. Its inhibitory effect on activation of trypsinogen by enteropeptidase, therefore, must be attributed to interaction with the zymogen rather than the enzyme. Kinetic measurements show that inhibition by rare earths is noncompetitive in nature. Binding of lanthanides to the tetraaspartyl sequence near the aminoterminus of trypsinogen may prevent this group from interacting with a critical specificity subsite on the enzyme.