Biochemical characterization of proline dehydrogenase in Arabidopsis mitochondria

• Published in: The FEBS journal 2014-06, Vol.281 (12), p.2794-2804
• Main Authors: Schertl, Peter, Cabassa, Cécile, Saadallah, Kaouthar, Bordenave, Marianne, Savouré, Arnould, Braun, Hans‐Peter
• Format: Article
• Language: English
• Subjects: Arabidopsis - enzymology; At3g30775; Biochemistry; Biosynthesis; Blotting, Western; Cells, Cultured; dehydrogenase; Electrophoresis, Polyacrylamide Gel; enzyme inhibitor; Enzymes; Glutamate Dehydrogenase - metabolism; Lactic Acid - pharmacology; Mitochondria; Mitochondria - enzymology; plant; Plant biology; proline; proline dehydrogenase (ProDH); Proline Oxidase - antagonists & inhibitors; Proline Oxidase - metabolism; Proteins; Stereoisomerism; stress response; Up-Regulation
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/febs.12821

Proline has multiple functions in plants. Besides being a building block for protein biosynthesis proline plays a central role in the plant stress response and in further cellular processes. Here, we report an analysis on the integration of proline dehydrogenase (ProDH) into mitochondrial metabolism in Arabidopsis thaliana. An experimental system to induce ProDH activity was established using cell cultures. Induction of ProDH was measured by novel photometric activity assays and by a ProDH in gel activity assay. Effects of increased ProDH activity on other mitochondrial enzymes were systematically investigated. Activities of the protein complexes of the respiratory chain were not significantly altered. In contrast, some mitochondrial dehydrogenases had markedly changed activities. Activity of glutamate dehydrogenase substantially increased, indicating upregulation of the entire proline catabolic pathway, which was confirmed by co‐expression analyses of the corresponding genes. Furthermore, activity of d‐lactate dehydrogenase was increased. d‐lactate was identified to be a competitive inhibitor of ProDH in plants. We suggest that induction of d‐lactate dehydrogenase activity allows rapid upregulation of ProDH activity during the short‐term stress response in plants. Proline protects plant cells under different stress conditions. The first enzyme of proline catabolism is proline dehydrogenase (ProDH). Here, we investigate effects of ProDH induction on mitochondrial metabolism. Besides enzymes of the proline catabolic pathway, D‐lactate dehydrogenase is induced. Lactate was identified as a competitive inhibitor of ProDH which might regulate ProDH during the plant stress response.