Abstract
Phosphoenolpyruvate carboxylase (EC 4.1.1.31; PEPCase) from Bryophyllum fedtschenkoi leaves has previously been shown to exist in two forms in vivo. During the night the enzyme is phosphorylated and relatively insensitive to feedback inhibition by malate whereas during the day the enzyme is dephosphorylated and more sensitive to inhibition by malate. These properties of PEPCase have now been investigated in leaves maintained under constant conditions of temperature and lighting. When leaves were maintained in continuous darkness and CO2-free air at 15°C, PEPCase exhibited a persistent circadian rhythm of interconversion between the two forms. There was a good correlation between periods during which the leaves were fixing respiratory CO2 and periods during which PEPCase was in the form normally observed at night. When leaves were maintained in continuous light and normal air at 15°C, starting at the end of a night or the end of a day, a circadian rhythm of net uptake of CO2 was observed. Only when these constant conditions were applied at the end of a day was a circadian rhythm of interconversions between the two forms of PEPCase observed and the rhythms of enzyme interconversion and CO2 uptake did not correlate in phase or period.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- CAM:
-
Crassulacean acid metabolism
- FW:
-
fresh weight
- PEPCase:
-
phosphoenolpyruvate carboxylase
- RuBPCase:
-
ribulose-1,5-bisphosphate carboxylase
References
Bollig, I.C., Wilkins, M.B. (1979) Inhibition of the circadian rhythm of CO2 metabolism in Bryophyllum leaves by cycloheximide and dinitrophenol. Planta 145, 105–112
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principle of protein-dye binding. Anal. Biochem. 72, 248–254
Brulfert, J., Queiroz, O. (1982) Photoperiodism and Crassulacean acid metabolism. III. Different characteristics of the photoperiod-sensitive and non-sensitive isoforms of phosphoenolpyruvate carboxylase and Crassulacean acid metabolism operation. Planta 184, 339–343
Brulfert, J., Vidal, J., Gadal, P., Queiroz, O. (1982) Daily rhythm of phosphoenolpyruvate carboxylase in Crassulacean acid metabolism plants. Immunological evidence for the absence of a rhythm in protein synthesis. Planta 156, 92–94
Brulfert, J., Vidal, J., Le Marechal, P., Gadal, P., Queiroz, O., Kluge, M., Kruger, I. (1986) Phosphorylation-dephosphorylation process as a probable mechanism for the diurnal regulatory changes of phosphoenolpyruvate carboxylase in CAM plants. Biochem. Biophys. Res. Commun. 136, 151–159
Buchanan-Bollig, I.C. (1984) Circadian rhythm in Kalanchoë: effects ot irradiance and temperature on gas exchange and carbon metabolism. Planta 160, 264–271
Buchanan-Bollig, I.C., Fischer, A., Kluge, M. (1984) Circadian rhythms in Kalanchoë: the pathway of 14CO2 fixation during prolonged light. Planta 161, 71–80
Buchanan-Bollig, I.C., Smith, J.A.C. (1984) Circadian rhythms in Crassulacean acid metabolism: phase relationships between gas exchange, leaf water relations and malate metabolism in Kalanchoë daigremontiana. Planta 161, 314–319
Fischer, A., Kluge, M. (1984) Studies on carbon flow in Crassulacean acid metabolism during the initial light period. Planta 160, 121–128
Greenway, H., Winter, K., Lüttge, U. (1978) Phosphoenolpyruvate carboxylase during development of Crassulacean acid metabolism and during a diurnal cycle in Mesembryanthemum crystallinum. J. Exp. Bot. 29, 547–559
Kenyon, W.H., Holaday, A.S., Black, C.C. (1981) Diurnal changes in metabolite levels and Crassulacean acid metabolism in Kalanchoë daigremontiana leaves. Plant Physiol. 68, 1002–1007
Kluge, M., Böcher, M., Jungnickel, G. (1980) Metabolic control of Crassulacean acid metabolism: evidence for diurnally changing sensitivity against inhibition by malate of PEP-carboxylase in Kalanchoë tubiflora Hamet. Z. Pflanzenphysiol. 97, 197–204
Kluge, M., Böhlke, C., Queiroz, O. (1981b) Crassulacean acid metabolism (CAM) in Kalanchoë: changes in intercellular CO2 concentration during a normal CAM cycle and during cycles in continuous light or darkness. Planta 152, 87–92
Kluge, M., Brulfert, J., Queiroz, O. (1981a) Diurnal changes in the regulatory properties of PEP-carboxylase in Crassulacean acid metabolism (CAM). Plant Cell Environ. 4, 251–256
Kluge, M., Ting, I.P. (1978) Crassulacean acid metabolism. Analysis of an ecological adaptation. Springer, Berlin Heidelberg New York
Littlejohn, R.O., Ku, M.S.B. (1984) Characterisation of early morning Crassulacean acid metabolism in Opuntia erinacea var. Columbiana (Griffiths) L. Benson. Plant Physiol. 74, 1050–1054
Manetas, Y. (1982) Changes in properties of PEP-carboxylase from the CAM plant Sedum prealtum DC upon dark/light transition and their stabilisation by glycerol. Photosynth. Res. 3, 321–333
Möllering, H. (1974) l-(-)-Malate. Determination with MDH and GOT. In: Methods of enzymatic analysis, 2nd edn., vol. 3, pp. 1589–1593, Bergmeyer, H.U., ed. Academic Press, New York
Nimmo, G.A., Nimmo, H.G., Fewson, C.A., Wilkins, M.B. (1984) Diurnal changes in the properties of phosphoenolpyruvate carboxylase in Bryophyllum leaves: a possible covalent modification. FEBS. Lett. 178, 199–203
Nimmo, G.A., Nimmo, H.G., Hamilton, I.D., Fewson, C.A., Wilkins, M.B. (1986) Purification of the phosphorylated night form and dephosphorylated day form of phosphoenol-pyruvate carboxylase from Bryophyllum fedtschenkoi. Biochem. J. 239, 213–220
Nungesser, D., Kluge, M., Tolle, H., Oppelt, W. (1984) A dynamic computer model of the metabolic and regulatory processes in Crassulacean acid metabolism. Planta 162, 204–214
Osmond, C.B. (1978) Crassulacean acid metabolism: a curiosity in context. Annu. Rev. Plant Physiol 29, 379–414
Osmond, C.B., Holtum, J.A.M. (1981) Crassulacean acid metabolism. In: The biochemistry of plants, vol. 8, pp. 283–328, Hatch, M.D., Boardman, N.K., eds. Academic Press, London New York
Ritz, D., Kluge, M., Veith, H.J. (1986) Mass-spectrometric evidence for the double-carboxylation pathway of malate synthesis by Crassulacean acid metabolism plants in light. Planta 167, 284–291
von Willert, D.J., Brinckmann, E., Scheitler, B., Thomas, D.A., Treichel, S. (1979) The activity and malate inhibition/stimulation of phosphoenolpyruvate carboxylase in Crassulacean acid metabolism plants in their natural environment. Planta 147, 31–36
Warren, D.M., Wilkins, M.B. (1961) An endogenous rhythm in the rate of dark-fixation of carbon dioxide in leaves of Bryophyllum fedtschenkoi. Nature 191, 686–688
Wilkins, M.B. (1959) An endogeneous rhythm in the rate of carbon dioxide output of Bryophyllum. I. Some preliminary experiments. J. Exp. Bot. 10, 377–390
Wilkins, M.B. (1960) An endogeneous rhythm in the rate of carbon dioxide output of Bryophyllum. II. The effects of light and darkness on the phase and period of the rhythm. J. Exp. Bot. 11, 269–288
Wilkins, M.B. (1962) An endogenous rhythm in the rate of carbon dioxide output of Bryophyllum. III. The effects of temperature on the phase and period of the rhythm. Proc. R. Soc. London B 156, 220–241
Wilkins, M.B. (1967) An endogenous rhythm in the rate of carbon dioxide output of Bryophyllum. IV. The dependence of rhythmicity on aerobic metabolism. Planta 72, 66–77
Wilkins, M.B. (1973) An endogenous cicadian rhythm in the rate of carbon dioxide output of Bryophyllum. VI. Action spectrum for the induction of phase shifts by visible radiation. J. Exp. Bot. 24, 488–496
Wilkins, M.B. (1983) The circadian rhythm of carbon-dioxide metabolism in Bryophyllum: the mechanism of phase-shift induction by thermal stimuli. Planta 157, 471–480
Wilkins, M.B. (1984) A rapid circadian rhythm of carbon-dioxide metabolism in Bryophyllum fedtschenkoi. Planta 161, 381–384
Wilkinson, M.J., Smith, H. (1976) Properties of phosphoenolpyruvate carboxylase from Bryophyllum fedtschenkoi leaves and fluctations in carboxylase activity during the endogenous rhythm of carbon dioxide output. Plant Sci. Lett. 6, 319–324
Winter, K. (1980a) Day/night changes in the sensitivity of phosphoenolpyruvate carboxylase to malate during Crassulacean acid metabolism. Plant Physiol. 65, 792–796
Winter, K. (1980b) Carbon dioxide and water vapour exchange in the Crassulacean acid metabolism plant Kalanchoë pinnata during a prolonged light period. Metabolic and stomatal control of carbon metabolism. Plant Physiol. 66, 917–921
Winter, K. (1982) Properties of phosphoenolpyruvate carboxylase in rapidly prepared, desalted leaf extracts of the Crassulacean acid metabolism plant Mesembryanthemum crystallimun L. Planta 154, 298–308
Author information
Authors and Affiliations
Additional information
To whom correspondence should be addressed.
Rights and permissions
About this article
Cite this article
Nimmo, G.A., Wilkins, M.B., Fewson, C.A. et al. Persistent circadian rhythms in the phosphorylation state of phosphoenolpyruvate carboxylase from Bryophyllum fedtschenkoi leaves and in its sensitivity to inhibition by malate. Planta 170, 408–415 (1987). https://doi.org/10.1007/BF00395034
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00395034