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Chromatographic and immunological evidence that chloroplastic and cytosolic pea (Pisum sativum L.) NADP-isocitrate dehydrogenases are distinct isoenzymes

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Abstract

Two NADP-isocitrate dehydrogenase isoenzymes designated as NADP-IDH1 and NADP-IDH2 (EC 1.1.1.42) were identified in pea (Pisum sativum) leaf extracts by diethylaminoethylcellulose chromatography. The predominant form was found to be NADP-IDH1 while NADP-IDH2 represented only about 4% of the total leaf enzyme activity. These enzymes share few common epitopes as NADP-IDH2 was poorly recognized by the specific polyclonal antibodies raised against NADP-IDH1, and as a consequence NADP-IDH2 does not result from a post-translational modification of NADP-IDH1. Subcellular fractionation and isolation of chloroplasts through a Percoll gradient, followed by the identification of the associated enzymes, showed that NADP-IDH1 is restricted to the cytosol and NADP-IDH2 to the chloroplasts. Compared with the cytosolic isoenzyme, NADP-IDH2 was more thermolabile and exhibited a lower optimum pH. The data reported in this paper constitute the first report that the chloroplastic NADP-IDH and the cytosolic NADP-IDH are two distinct isoenzymes. The possible functions of the two isoenzymes are discussed.

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Abbreviations

BSA:

bovine serum albumin

DEAE:

diethylaminoethyl

NADP-IDH:

NADP-isocitrate dehydrogenase

NADP-IDH1 :

cytosolic NADP-IDH

NADP-IDH2 :

chloroplastic NADP-IDH

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Authors and Affiliations

  1. Physiologie Végétale Moléculaire URA CNRS D 1128, Université Paris Sud, Bâtiment 430, F-91405, Orsay Cedex, France

    Ri-Dong Chen & Pierre Gadal

  2. Photosynthèse et Métabolisme, URA CNRS D 1128, Université Paris Sud, Bâtiment 430, F-91405, Orsay Cedex, France

    Evelyne Bismuth & Marie-Louise Champigny

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  1. Ri-Dong Chen
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  2. Evelyne Bismuth
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  3. Marie-Louise Champigny
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  4. Pierre Gadal
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Chen, RD., Bismuth, E., Champigny, ML. et al. Chromatographic and immunological evidence that chloroplastic and cytosolic pea (Pisum sativum L.) NADP-isocitrate dehydrogenases are distinct isoenzymes. Planta 178, 157–163 (1989). https://doi.org/10.1007/BF00393190

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  • DOI: https://doi.org/10.1007/BF00393190

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