Abstract
NADP-malate dehydrogenase extracted from darkened leaves of the C3 plants pea, barley, wheat and spinach was activated by reduced glutathione, a monothiol, as well as by dithiothreitol (DTT). However, in the C4 plants maize and Flaveria trinervia, only dithiothreitol could effectively activate the enzyme. There was no activation of the maize enzyme and little or no activation of the F. trinervia enzyme by glutathione. The failure of glutathione to activate NADP-MDH in leaf extracts of maize and F. trinervia may indicate there is some difference in disulfide groups of the protein compared to the C3 plant enzyme. Both DTT and glutathione could activate NADP-malate dehydrogenase in a partially purified enzyme preparation from pea leaves with or without addition of partially purified thioredoxin. However, the required concentration of reductant was lower with addition of thioredoxin than in its absence. In extracts of C3 species and the partially purified pea enzyme the level of activation after 40 to 60 min under aerobic conditions was higher (up to twofold) with DTT than with glutathione. Under anaerobic conditions, the initial rate of activation was about twice as high with DTT as with glutathione, but the total activation after 40 to 60 min was similar. Ascorbate was totally ineffective as a reducing agent in activating NADP-MDH from C3 or C4 plants, possibly due to its more positive redox potential.
Similar content being viewed by others
Abbreviations
- Chl:
-
Chlorophyll
- DTT:
-
Dithiothreitol
- GSH:
-
Reduced Glutathione
- NADP-MDH:
-
NADP-malate Dehydrogenase
References
Anderson, LE, Ashton, AR, Mohamed, AH and Scheibe, R (1982) Light/dark modulation of enzyme activity in photosynthesis. Bioscience 32: 103–107
Bradford, MM (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein dye binding. Anal Biochem 72: 248–254
Buchanan, BB (1980) Role of light in the regulation of chloroplast enzymes. Annu Rev Plant Physiol 312: 341–374
Buchanan, BB, Schurmann, P and Kalberer, PP (1971) Ferredoxin-activated fructose diphosphatase of spinach chloroplasts. J Biol Chem 246: 5952–5959
Edwards, GE, Nakamoto, H, Burnell, JN and Hatch, MD (1985) Pyruvate, Pi dikinase and NADP-malate dehydrogenase in C4 photosynthesis: Properties and mechanism of light/dark regulation. Annu Rev Plant Physiol 36: 255–286
Ferte, N, Jacquot, J-P and Meunier, J-C (1986) Structural, immunological and kinetic comparisons of NADP-dependent malate dehydrogenases from spinach (C3) and corn (C4) chloroplasts. Eur J Biochem 154: 587–594
Fickenscher, K and Scheibe, R (1983) Purification and properties of NADP-dependent malate dehydrogenase from pea leaves. Biochem Biophys Acta 749: 249–254
Foyer, C and Halliwell, B (1976) The presence of glutathione and glutathione reductase in chloroplasts: A proposed role in ascorbic acid metabolism. Planta 133: 21–25
Holmgren, A (1977) Bovine thioredoxin system. J Biol Chem 252: 4600–4606
Jenkins, CLD, Anderson and Hatch, MD (1986) NADP-malate dehydrogenase from Zea mays leaves: Amino acid comnposition and thio content of active and inactive forms. Plant Sci Lett 45: 1–7
Jocelyn, PC (1972) Biochemistry of the Thiol Group. New York: Academic Press
Nakamoto, H and Edwards, GE (1983) Influence of oxygen and temperature on the dark inactivation of pyruvate, Pi dikinase and NADP-malate dehydrogenase in maize. Plant Physiol 71: 568–573
Nakano, Y and Asada, K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22: 867–880
Rebeille, F and Hatch, MD (1986) Regulation of NADP-malate dehydrogenase in C4 plants: Effect of varying NADPH to NADP rations and thioredoxin redox state on enzyme activity in reconstituted systems. Arch Biochem Biophys 249: 164–170
Reid, EE (1958) Organic Chemistry of Bivalent Sulfur. Vol 1, New York: Chemical Publishing Co
Scheibe, R and Anderson, LE (1981) Dark modulation of NADP-dependent malate dehydrogenase and glucose-6-phosphate dehydrogenase in the chloroplast. Biochim Biophys Acta 636: 58–64
Scheibe, R and Fickenscher, K (1984) In: Sybesma, C (ed.) Advances in Photosynthesis Research, Vol 2, pp. 529–532. The Hague: Martinus Nijhoff/Dr W Junk Publishers
Scheibe, R, Fickenschen, K and Ashton, AR (1986) Studies on the mechanism of the reductive activation of NADP-malate dehydrogenase by thioredoxin. m and low-molecular weight thiols. Biochim Biophys Acta 870: 191–197
Schurmann, P, Wolosiuk, RA, Breazeale, VD and Buchanan, BB (1976) Two proteins function in the regulation of photosynthetic CO2 assimilation in chloroplasts. Nature 263: 257–258
Vivekanandan M and Edwards GE (1987) Light and dark anaerobic activation of NADP-malate dehydrogenase in pea leaves and chloroplasts. Proc VII Int Congress Photosynthesis, Progress in Photosynthesis Res, J Biggins (ed.) Vol 3, pp. 257–260.
Wintermans, JFGM and De, Mots, A (1965) Spectrotometric characteristics of chlorophyll a and b and their pheophytins in ethanol. Biochim Biophys Acta 109: 448–453
Wolosiuk, RA and Buchanan, BB (1977) Thioredoxin and glutathione regulate photosynthesis in chloroplasts. Nature 266: 565–567
Wolosiuk, RA, Buchanan, BB and Crawford, NA (1977) Regulation of NADP-malate dehydrogenase by the light-activated ferredoxin/thioredoxin system of chloroplasts. FEBS Lett 81: 253–258
Wolosiuk RA, Schurmann P and Buchanan BB (1980) Thioredoxin and ferredoxinthioredoxin reductase of spinach chloroplasts. In: San Pietro A (ed.) Photosynthesis and Nitrogen Fixation. 69: 382–391. Series: Colowick SP and Kaplan NO (eds), Methods in Enzymology
Young, LCT and Conn, EE (1956) The reduction and oxidation of glutathione by plant mitochondria. Plant Physiol 31: 205–211
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Vivekanandan, M., Edwards, G.E. Activation of NADP-malate dehydrogenase in C3 plants by reduced glutathione. Photosynth Res 14, 113–124 (1987). https://doi.org/10.1007/BF00032316
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00032316