Abstract
Adenylate levels and oxygen evolution in isolated intact spinach chloroplasts were measured in the presence of nitrite or oxaloacetate, which do not consume ATP during reduction, and of bicarbonate and glycerate the reduction of which requires ATP. The following results were obtained:
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1.
In chloroplasts incubated in the dark with nitrite, oxaloacetate or bicarbonate, the concentration of AMP was high and that of ATP low. The concentration of ADP was usually but not always lower than that of AMP. Light induced a rapid increase in the level of ATP which was accompained by a marked decrease in AMP and a slight decrease in ADP. The ratio of ATP/ADP rarely increased above 3 in the light, even in the absence of ATP consumption. Glycerate differed from other substrates in that it drastically decreased the chloroplast ATP level in the dark and kept its concentration low, even under illumination. The rate of glycerate-dependent oxygen evolution was high.
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2.
Mass action ratios (ATP) (AMP)/(ADP)2 were close to 0.5 in the dark and in the presence of various substrates, except glycerate, indicating that adenylates were close to adenylate kinase equilibrium. When CO2 served as the substrate, (ATP) (AMP)/(ADP)2 values were often higher than 0.5 in the light. In the presence of glycerate, the values were always much below 0.5.
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3.
The energy charge [2(ATP)+(ADP)]/2[(ATP) +(ADP)+(AMP)] of the chloroplast adenylate system was between 0.27 and 0.6 in the dark. It increased, but scarcely exceeded 0.8, in the light. Energy charge was usually higher with bicarbonate than with phosphoglycerate, and always low with glycerate, both in the dark and in the light.
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Abbreviations
- E.C.:
-
energy charge
- ATP:
-
adenosine triphosphate
- ADP:
-
adenosine diphosphate
- AMP:
-
adenosine monophosphate
- GTP:
-
guanosine triphosphate
- MES:
-
2(N-morpholino)-ethanesulfonic acid
- HEPES:
-
N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid
- CCCP:
-
carbonylcyanide-m-chlorophenylhydrazone
References
Atkinson, D.E., Walton, G.M.: Adenosine triphosphate conservation in metabolic regulation. J. Biol. Chem. 242, 3239–3241 (1967)
Atkinson, D.E.: The energy charge of the adenylate pool as a regulatory parameter. Interaction with feedback modifiers. Biochemistry 7, 4030–4034 (1968)
Bomsel, J., Pradet, A.: Study of adenosine 5′-mono-, di- and triphosphates in plant tissues. IV. Regulation of the level of nucleotides, in vivo, by adenylate kinase; Theoretical and experimental study. Biochim. Biophys. Acta 162, 230–242 (1968)
Bowen, W.J., Kerwin, T.D.: The kinetics of myokinase. II. Studies of heat denaturation, the effects of salts and the state of equilibrium. Arch. Biochem. Biophys. 64, 278–284 (1956)
Chance, B., Williams, G.R.: A method for the localization of sites for oxidative phosphorylation. Nature 176, 250–254 (1955)
Eggleston, L.V., Hems, R.: Separation of adenosine phosphates by paper chromatography and the equilibrium constant of the myokinase system. Biochem. J. 52, 156–160 (1952)
Green, I., Brown, J.R.C., Mommaerts, W.F.H.M.: Adenosine triphosphatase systems of muscle. II. Identification of the reaction products. J. Biol. Chem. 205, 493–501 (1953)
Heber, U., Santarius, K.A.: Direct and indirect transfer of ATP and ADP across the chloroplast envelope. Z. Naturforsch. 25b. 718–728 (1970)
Heber, U., Kirk, M.R., Gimmler, H., Schäfer, G.: Uptake and reduction of glycerate by isolated chloroplasts. Planta 120, 31–46 (1974)
Heber, U., Kirk, M.R.: Flexibility of coupling and stoichiometry of ATP formation in intact chloroplasts. Biochim. Biophys. Acta 376, 136–150 (1975)
Heldt, H.W., Schwalbach, K.: The participation of GTP-AMP-P transferase in substrate level phosphate transfer of rat liver mitochondria. Eur. J. Biochem. 1, 199–206 (1967)
Heldt, H.W.: Adenine nucleotide translocation in spinach chloroplasts. FEBS Lett. 5, 11–14 (1969)
Heldt, H.W., Sauer, F.: The inner membrane of the chloroplast envelope as the site of specific metabolite transport. Biochim. Biophys. Acta. 234, 83–91 (1971)
Heldt, H.W.: Metabolite transport in intact chloroplasts. In: The Intact Chloroplast, pp. 215–234, Barber, J., ed. Elsevier: Amsterdam 1976
Heldt, H.W., Chon, C.J., Maronde, D., Herold, A., Stankovic, Z.S., Walker, D.A., Kraminer, A., Kirk, M.R., Heber, U.: Role of orthophosphate and other factors in the regulation of starch formation in leaves and isolated chloroplasts. Plant Physiol. 59, 1146–1155 (1977)
Inoue, Y., Kobayashi, Y., Shibata, K., Heber, U.: Synthesis and hydrolysis of ATP by intact chloroplasts under flash illumination and in darkness. Biochim. Biophys. Acta 504, 142–152 (1978)
Jensen, R.G., Bassham, J.A.: Photosynthesis by isolated chloroplasts. Proc. Natl. Acad. Sci. USA 56, 1095–1101 (1966)
Klingenberg, M., Heldt, H.W., Pfaff, E.: In: The energy level and metabolic control in mitochondria, pp. 237–253, Papa, S., Tager, M., Quagliariello, E., Slater, E.C., eds. Bari: Adriatica Editrice 1969
Kobayashi, Y., Inoue, Y., Shibata, K., Heber, U.: Control of electron flow in intact chloroplasts by the intrathylakoid pH, not by the phosphorylation potential. Planta 146, 481–486 (1979)
Kraayenhof, R.: “State 3-State 4 transition” and phosphate potential in “Class I” spinach chloroplasts. Biochim. Biophys. Acta 180, 213–215 (1969)
Krause, G.H.: Light-induced movement of magnesium ions in intact chloroplasts — spectroscopic determination with eriochrome blue SE. Biochim. Biophys. Acta 460, 500–501 (1977)
Lilley, R., Chon, C.J., Mosbach, A., Heldt, H.W.: The distribution of metabolites between spinach chloroplasts and medium during photosynthesis in vitro. Biochim. Biophys. Acta 460, 259–272 (1977)
Pfaff, E., Klingenberg, M., Ritt, E., Vogell, W.: Korrelation des unspezifisch permeablen mitochondrialen Raumes mit dem “Intramembran-Raum”. Eur. J. Biochem. 5, 222–232 (1968)
Purczeld, P., Chon, C.J., Portis Jr., A.R., Heldt, H.W., Heber, U.: The mechanism of the control of carbon fixation by the pH in the chloroplast stroma. Studies with nitrite-mediated proton transfer across the envelope. Biochim. Biophys. Acta 501, 488–498 (1978)
Santarius, K.A., Heber, U.: Changes in the intracellular levels of ATP, ADP, AMP and Pi and regulatory function of the adenylate system in leaf cells during photosynthesis. Biochim. Biophys. Acta 102, 39–54 (1965)
Walker, D.A.: Plastid and intracellular transport. In: Encyclopedia of Plant Physiol., New Series, Transport in Plants III, pp. 85–136, Stocking, C.R., Heber, U., eds. Berlin, Heidelberg, New York: Springer 1976
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Kobayashi, Y., Inoue, Y., Furuya, F. et al. Regulation of adenylate levels in intact spinach chloroplasts. Planta 147, 69–75 (1979). https://doi.org/10.1007/BF00384593
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DOI: https://doi.org/10.1007/BF00384593