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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- FPLC :
-
Fast protein liquid chromatography
References
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of proteins utilizing the proteindye binding. Anal Biochem 72:248–254
Cerff R (1982) Separation and purification of NAD- and NADP-linked glyceraldehyde-3-phosphate dehydrogenases from higher plants. In: Edelman M, Hallik RB, Chua NH (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, Amsterdam, pp 683–694
Fotherhill-Gilmore LA, Michels PAM (1993) The evolution of glycolysis. Prog Biophys Mol Biol 59:105–235
Haselkorn R, Buikema WJ (1992) Nitrogen fixation by cyanobacteria. In: Stacey G, Burris RH, Evans HJ (eds) Biological nitrogen fixation. Chapman & Hall, New York London, pp 166–190
Hood W, Carr NG (1967) A single glyceraldehyde-3-phosphate dehydrogenase active with NAD and NADP in Anabaena variabilis. Biochem Biophys Acta 146:309–311
Hood W, Carr NG (1969) Association of NAD and NADP linked glyceraldehyde-3-phosphate dehydrogenase in the blue-green alga Anabaena variabilis. Planta 80:250–258
Jakowitsch J, Bayer MG, Maier T, Lüttke A, Gebhart UB, Brandtner M, Hamilton B, Neumann-Spallart C, Michailowski CB, Bohnert HJ, Schenk HEA, Löffelhardt W (1993) Sequence analysis of pre-ferredoxin-NADP-reductase cDNA from Cyanophora paradoxa specifying a precursor for a nucleus-encoded cyanelle polypeptide. Plant Mol Biol 21:1023–1033
Kremer BP, Kies L, Rostani-Rabet A (1979) Photosynthetic performance of cyanelles in the endocyanomes Cyanophora, Glaucosphaera, Gloeochaete, and Glaucocystis. Z Pflanzenphysiol 92:303–317
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227:680–685
Liaud MF, Valentin C, Brandt U, Bouget FY, Kloareg B, Cerff R (1993) The GAPDH gene system of the red alga Chondrus crispus: promotor structures, intron/exon organization, genomic complexity and differential expression of genes. Plant Mol Biol 23:981–994
Martin W, Brinkmann H, Savonna C, Cerff R (1993) Evidence for a chimeric nature of nuclear genomes: eubacterial origin of cukaryotic glyceraldehyde-3-phosphate dehydrogenase genes. Proc Natl Acad USA 90:8692–8696
Mateos MI, Serrano A (1992) Occurrence of phosphorylating and non-phosphorylating NADP-dependent glyceraldehyde-3-phosphate dehydrogenases in photosynthetic organisms. Plant Sci 84:163–170
McFadden BY, Shively JM (1991) Bacterial assimilation of carbon dioxide by the Calvin cycle. In: Shively JM, Barton LL (eds) Variations of autotrophic life. Academic Press, London, pp 25–49
Mucke H, Löffelhardt W, Bohnert HJ (1980) Partial characterization of the genome of the “endosymbiotic” cyanelles from Cyanophora paradoxa. FEBS Lett 111:347–352
Pearce J, Carr NG (1969) The incorportation and metabolism of glucose by Anabaena variabilis. J Gen Microbiol 54:451–462
Pelroy RA, Rippka R, Stanier RY (1972) Metabolism of glucose by unicellular blue-green algae. Arch Mikrobiol 87:303–332
Schenk HEA (1992) Cyanobacterial symbioses. In: Ballows A, Trüper HG, Dworkin M, Harder W, Schleifer KH (eds) The prokaryotes, vol 4. Springer, New York, pp 3819–3854
Schlichting R, Zimmer W, Bothe H (1990) Exchange of metabolites in Cyanophora paradoxa and its cyanelles. Bot Acta 103: 392–398
Schrautemeier B, Böhme H, Böger P (1984) In vitro studies on pathways and regulation of electron transport to nitrogenase with a cell-free extract from heterocysts of Anabaena variabilis. Arch Microbiol 137:14–20
Serrano A, Llobell A (1993) Occurrence of two isoforms of glutathione reductase in the gree alga Chlamydomonas reinhardtii. Planta 190:199–205
Trench RK (1982) Cyanelles. In: Schiff JA, Lyman H (eds) On the origin of chloroplasts. Elsevier/North-Holland, New York, pp 55–76
Udvardy J, Balogh Á, Farkas GL (1982) Modulation of glyceraldehyde-3-phosphate dehydrogenase in Anacystis nidulans by glutathione. Arch Microbiol 133:2–5
Wasmann CC, Löffelhardt W, Bohnert HJ (1987) Cyanelles: organization and molecular biology. In: Fay P, Van Baalen C (eds) The cyanobacteria. Elsevier, Amsterdam, pp 301–324
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Serrano, A., Löffelhardt, W. Identification of two different glyceraldehyde-3-phosphate dehydrogenases (phosphorylating) in the photosynthetic protist Cyanophora paradoxa . Arch. Microbiol. 162, 14–19 (1994). https://doi.org/10.1007/BF00264367
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00264367