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Intrasteric inhibition in redox signalling: light activation of NADP-malate dehydrogenase

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Abstract

Chloroplast NADP-dependent malate dehydrogenase (NADP-MDH, EC 1.1.1.82) is inactive in the dark and activated in the light via a reduction of specific disulfides by thiol-disulfide interchange with thioredoxin, reduced by the photosynthetic electron transfer. Compared to the constitutively active NAD-dependent forms, NADP-MDH exhibits two regulatory disulfides per subunit, one located in an N-terminal extension and the other in a C-terminal extension. Convergent information gathered from biochemical, site-directed mutagenesis and structural approaches allowed to solve almost completely the activation mechanism. In the oxidized enzyme, the C-terminal extension is pulled back by the disulfide bridge toward the active-site cleft where the penultimate C-terminal glutamate interacts with one of the arginines involved in substrate binding, thus acting as an internal inhibitor obstructing the access of oxaloacetate. The N-terminal extensions are located at the subunit interface area and rigidify the overall structure of the dimer. Their reduction by reduced thioredoxin triggers a conformational change of the active site towards high-activity conformation, whereas the reduction of the C-terminal bridge expells the C-terminal end from the active site, thus opening the way for the substrate.

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  1. Institut de Biotechnologie des Plantes, UMR CNRS, Université de Paris-Sud, 8618, Bât. 630, 91405, Orsay Cedex, France

    Myroslawa Miginiac-Maslow

  2. Laboratoire de RMN Biomoléculaire, UMR CNRS, Université Claude Bernard Lyon I and Ecole Supérieure de Chimie, Physique et Electronique de Lyon, Villeurbanne, France

    Jean-Marc Lancelin

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  1. Myroslawa Miginiac-Maslow
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  2. Jean-Marc Lancelin
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Correspondence to Myroslawa Miginiac-Maslow.

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Miginiac-Maslow, M., Lancelin, JM. Intrasteric inhibition in redox signalling: light activation of NADP-malate dehydrogenase. Photosynthesis Research 72, 1–12 (2002). https://doi.org/10.1023/A:1016099228450

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