At5g50600 encodes a member of the short-chain dehydrogenase reductase superfamily with 11 β- and 17 β-hydroxy steroid dehydrogenase activities associated with Arabidopsis thaliana seed oil bodies

• Published in: Biochimie Vol. 89; no. 2; pp. 222 - 229
• Main Authors: d'Andrea, Sabine, Canonge, Michel, Beopoulos, Athanastos, Jolivet, Pascale, Hartmann, Marie-Andrée, Miquel, Martine, Lepiniec, Loic, Chardot, Thierry
• Format: Journal Article
• Language: English
• Published: Elsevier 01.02.2007
• Subjects: Chemical and Process Engineering; Engineering Sciences; Food engineering; Life Sciences; Arabidopsis thaliana; hydroxysteroid dehydrogenase reductase; oil bodies; short-chain dehydrogenase reductase; estradiol oxidation
• ISSN: 0300-9084; 1638-6183
• DOI: 10.1016/j.biochi.2006.09.013

In a previous work, we presented evidence for the presence of a protein encoded by At5g50600 in oil bodies (OBs) from Arabidopsis thaliana [P. Jolivet, E. Roux, S. D'Andrea, M. Davanture, L. Negroni, M. Zivy, T. Charclot, Protein composition of oil bodies in Arabidopsis thaliana lecotype WS, Plant Physiol. Biochem. 42 (2004) 501-509]. Using specific antibodies and proteomi c techniques, we presently confirm the existence of this protein, which is a member of the short-chain steroid dehydrogenase reductase superfamily. We have measured its activity toward various steroids (cholesterol, dehydroepiandrosterone, cortisol, corticosterone, estradiol, estrone) and NAD(P)(H), either within purified OBs or as a purified bacterially expressed chimera. Both enzymatic systems (OBs purified from A. thaliana seeds as well as the chimeric enzyme) exhibited hydroxysteroid dehydrogenase (HSD) activity toward estradiol (17 beta-hydroxysteroid) with NAD+ or NADP(+) NADP+ being the preferred cofactor. Low levels of activity were observed with cortisol or corticosterone (Ill3-hydroxysteroids), but neither cholesterol nor DHEA (3 beta-hydroxysteroids) were substrates, whatever the cofactor used. Similar activity profiles were found for both enzyme sources. Purified OBs were found to be also able to catalyze estrone reduction (17 beta-ketosteroid reductase activity) with NADPH. The enzyme occurring in A. thaliana OBs can be classified as a NADP+-dependent 11 beta,17 beta-hydroxysteroid dehydrogenase/17 beta-ketosteroid reductase. This enzyme probably corresponds to AtHSD1, which is encoded by At5g50600. However, its physiological role and substrates still remain to be determined.