NADP+-dependent cytosolic isocitrate dehydrogenase provides NADPH in the presence of cadmium due to the moderate chelating effect of glutathione

Abstract

Cadmium (Cd²⁺) is toxic to living organisms because it causes the malfunction of essential proteins and induces oxidative stress. NADP⁺-dependent cytosolic isocitrate dehydrogenase (IDH) provides reducing energy to counteract oxidative stress via oxidative decarboxylation of isocitrate. Intriguingly, the effects of Cd²⁺ on the activity of IDH are both positive and negative, and to understand the molecular basis, we determined the crystal structure of NADP⁺-dependent cytosolic IDH in the presence of Cd²⁺. The structure includes two Cd²⁺ ions, one coordinated by active site residues and another near a cysteine residue. Cd²⁺ presumably inactivates IDH due to its high affinity for thiols, leading to a covalent enzyme modification. However, Cd²⁺ also activates IDH by providing a divalent cation required for catalytic activity. Inactivation of IDH by Cd²⁺ is less effective when the enzyme is activated with Cd²⁺ than Mg²⁺. Although reducing agents cannot restore activity following inactivation by Cd²⁺, they can maintain IDH activity by chelating Cd²⁺. Glutathione, a cellular sulphydryl reductant, has a moderate affinity for Cd²⁺, allowing IDH to be activated with residual Cd²⁺, unlike dithiothreitol, which has a much higher affinity. In the presence of Cd²⁺-consuming cellular antioxidants, cells must continually supply reductants to protect against oxidative stress. The ability of IDH to utilise Cd²⁺ to generate NADPH could allow cells to protect themselves against Cd²⁺.