Identification of two different glyceraldehyde-3-phosphate dehydrogenases (phosphorylating) in the photosynthetic protist Cyanophora paradoxa |
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Authors: | Aurelio Serrano Wolfgang Löffelhardt |
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Institution: | (1) Instituto de Bioquímica Vegetal y Fotosíntesis, CSIC y Universidad de Sevilla, Apdo. 1113, E-41080 Sevilla, Spain;(2) Institut für Biochemie und Molekulare Zellbiologie der Universität Wien, A-1030 Vienna, Austria;(3) Ludwig Boltzmann-Forschungsstelle für Biochemie, A-1030 Vienna, Austria |
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Abstract: | Two different glyceraldehyde-3-phosphate (G3P) dehydrogenase (phosphorylating) activities, namely NAD- and NADP-dependent, have been found in cell extracts of the cyanelle-bearing photosynthetic protist Cyanophora paradoxa. Whereas the two G3P dehydrogenase activities were detected with similar specific activity levels (0.1 to 0.2 U/mg of protein) in extracts of the photosynthetic organelles (cyanelles), only the NAD-dependent activity was found in the cytosol. Thus, a differential intracellular localization occurred. The perfect overlapping of the two G3P dehydrogenase activity peaks of the cyanelle in both hydrophobic interaction chromatography and subsequent FPLC (fast protein liquid chromatography) gel filtration indicated that the two activities were due in fact to a single NAD(P)-dependent G3P dehydrogenase (EC 1.2.1.-) with a molecular mass of 148,000. SDS-PAGE of active fractions from FPLC gel filtration showed that the intensity of the major protein band (molecular mass, 38,000) of the enzyme preparation clearly paralleled the activity elution profile, thus suggesting a tetrameric structure for the cyanelle dehydrogenase. On the other hand, FPLC gel filtration analysis of the cytoplasmic fraction revealed a NAD-dependent G3P dehydrogenase with a native molecular mass of 142,000, being equivalent to the classical glycolytic enzyme (EC 1.2.1.12) present in the cytosol of all the organisms so far studied. The significance of these results is discussed taking into account that the cyanobacteria, photosynthetic prokaryotes which share many structural and biochemical features with cyanelles and are considered as their ancestors, have a similar NAD(P)-dependent G3P dehydrogenase.Abbreviation
FPLC
Fast protein liquid chromatography |
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Keywords: | Cyanophora paradoxa NAD(P)-dependent glyceraldehyde-3-phosphate dehydrogenase NAD-dependent glyceraldehyde-3-phosphate dehydrogenase Cyanelles |
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