Interaction of analogues of substrate with NADP-malic enzyme from maize leaves

Abstract

The effect of structural analogues of l-malate was studied on NADP-malic enzyme purified from Zea mays L. leaves. Among the compounds tested, the organic acids behaved as more potent inhibitors at pH 7.0 than at pH 8.0, suggesting that the dimeric form was more susceptible to the inhibition than the tetrameric form of the enzyme.

Oxalate, ketomalonate, hydroxymalonate, malonate, oxaloacetate, tartrate, α-hydroxybutyrate, α-ketobutyrate, α-ketoglutarate and α-hydroxyglutarate exhibited linear competitive inhibition with respect to the substrate l-malate at pH 8.0. On the other hand, glyoxylate and glycolate turned out to be non-competitive inhibitors, while glycolaldehyde, succinate, fumarate, maleate and β- and γ-hydroxybutyrate had no effect on the enzyme activity, at the concentrations assayed. These results suggest that the extent of inhibition was dependent on the size of the analogues and that the presence of an 1-carboxyl group along with a 2-hydroxyl or 2-keto group was important for binding of the substrate analogue to the enzyme.