Citation

Clemens F, Drutkowski G, Wiese M, Frohberg P. Biochim. Biophys. Acta 2001; 1549(1): 88-98.

Copyright

(Copyright © 2001, Elsevier Publishing)

DOI

10.1016/s0167-4838(01)00248-5

PMID

11566371

Abstract

Several compounds containing an amidrazone moiety are known to be potent inhibitors of lipoxygenase-1 activity from soybeans (L-1) with IC(50)-values in the range of 10 microM to 38 nM. Recently it was proposed that phenylhydrazones act as irreversible mechanism-based inhibitors of lipoxygenases. Because of the structural similarities between both compounds it was assumed for the amidrazones to affect the lipoxygenase reaction in the same suicide manner. Cyclisation of the amidrazone moiety to the corresponding triazoline should yield compounds without substrate properties. However, they are still able to inactivate the enzyme. The inhibition of L-1 from soybeans by two representative compounds of a series of amidrazones and triazolines has been characterised as a slow, tight-binding interaction via a two-step mechanism. Dialysis experiments indicate the reversible nature of interaction of the amidrazone with the ferrous enzyme while the ferric enzyme was irreversibly inactivated. In contrast, the interaction of the triazoline with both the ferric and ferrous species of the enzyme was completely reversible which demonstrates the noncovalent and reversible mode of binding and inactivation. The triazoline was found not to be a substrate of the dioxygenase reaction of lipoxygenase whereas the amidrazone is only a very poor substrate of the enzymatic oxidation reaction. The presented results point out the inhibition of L-1 by amidrazones and triazolines to fall into the same kinetic classification. Therefore it is obvious that the inhibition of L-1 by these compounds cannot be attributed to a truly mechanism-based inactivation.

Language: en

Keywords

Glycine max; Hydrazones; Kinetics; Lipoxygenase; Lipoxygenase Inhibitors; Models, Chemical; Molecular Structure; Structure-Activity Relationship; Triazoles