Planta

, Volume 168, Issue 4, pp 536–545 | Cite as

Control of CO2 fixation regulation of stromal fructose-1,6-bisphosphatase in spinach by pH and Mg2+ concentration

  • A. Gardemann
  • D. Schimkat
  • H. W. Heldt
Article
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Accepted:

Abstract

The effect of pH and of Mg2+ concentration on the light activated form of stromal fructose-1,6-bisphosphatase (FBPase) was studied using the enzyme rapidly extracted from illuminated spinach chloroplasts. The (fructose-1,6-bisphosphate4-)(Mg2+) complex has been identified as the substrate of the enzyme. Therefore, changes of pH and Mg2+ concentrations have an immediate effect on the activity of FBPase by shifting the pH and Mg2+ dependent equilibrium concentration of the substrate. In addition, changes of pH and Mg2+ concentration in the assay medium have a delayed effect on FBPase activity. A correlation of the activities observed using different pH and Mg2+ concentrations indicates, that the effect is not a consequence of the pH and Mg2+ concentration as such, but is caused by a shift in the equilibrium concentration of a hypothetical inhibitor fructose-1,6-bisphosphate3- (uncomplexed), resulting in a change of the activation state of the enzyme. The interplay between a rapid effect on the concentration of the substrate and a delayed effect on the activation state enables a rigid control of stromal FBPase by stromal Mg2+ concentrations and pH. Fructose-1,6-bisphosphatase is allosterically inhibited by fructose-6-phosphate in a sigmoidal fashion, allowing a fine control of the enzyme by its product.

Key words

CO2 fixation Fructose-1,6-bisphosphatase Photosynthesis Regulation (fructose-1,6-bisphosphatase) Spinacia (chloroplasts) 

Abbreviations

Fru1,6 bis P

fructose-1,6-bisphosphate

Fru6P

fructose-6-phosphate

FBPase

fructose-1,6-bisphosphatase

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References

  1. Baier, D., Latzko, E. (1975) Properties and regulation of C-1-fructose-1,6-diphosphatase from spinach chloroplasts. Biochim. Biophys. Acta 396, 141–148Google Scholar
  2. Buchanan, B.B. (1980) Role of light in the regulation of chloroplast enzymes. Annu. Rev. Plant Physiol. 31, 341–374Google Scholar
  3. Charles, S.A., Halliwell, B. (1980) Properties of freshly purified and thiol-treated spinach chloroplast fructose bisphosphatase. Biochem. J. 185, 689–693Google Scholar
  4. Falkner, G., Horner, F., Werdan, K., Heldt, H.W. (1976) pH Changes in the cytoplasm of the blue-green alga Anacystis nidulans caused by light-dependent proton flux into the thylakoid space. Plant Physiol. 58, 717–718Google Scholar
  5. Gontero, B., Meunier, J.-C., Buc, J., Richard, J. (1984) The slow pH induced conformational transition of chloroplast fructose-1,6-bisphosphatase and the control of the Calvin cycle. Eur. J. Biochem. 145, 485–488Google Scholar
  6. Heldt, H.W., Chon, C.J., Lilley, R.McC. Portis, A.R. (1978) The role of fructose and sedoheptulosebisphosphatase in the control of CO2 fixation. Evidence from the effects of Mg2+ concentration, pH and H2O2. Proc. 4th Int. Congr. on Photosynthesis, pp. 469–478, Hall, D.O., Coombs, J., Goodwin, T.W., eds. The Biochemical Society, LondonGoogle Scholar
  7. Heldt, H.W., Gardemann, A., Gerhardt, R., Herzog, B., Stitt, M., Wirtz, W. (1984) The regulation of CO2 fixation and of sucrose synthesis in plants. Proc. 6th Int. Congr. on Photosynthesis, vol 3, pp. 617–624, Sybesma C., ed. Nijhoff-Junk, The HagueGoogle Scholar
  8. Itaya, K., Ui, M. (1966) A new micromethod for the colorimetric determination of inorganic phosphate. Clin. Chim. Acta 14, 361–366Google Scholar
  9. Krause, G.H. (1977) Light-induced movement of magnesium ions in intact chloroplasts. Spectroscopic determination with Eriochrome blue S. E. Biochim. Biophys. Acta 460, 500–510Google Scholar
  10. Laing, W.A., Stitt, M., Heldt, H.W. (1981) Control of CO2-fixation: Changes in the activity of ribulose-5-phosphate kinase and fructose- and sedoheptulose-bisphosphatase in chloroplasts. Biochim. Biophys. Acta 637, 348–359Google Scholar
  11. Leegood, R.C. (1981) Measurement of fructose bisphosphatase activity from intact chloroplasts. Report of the Research of A.R.C. Research Group on Photosynthesis, pp. 26–32, Department of Botany, Sheffield, UKGoogle Scholar
  12. Leegood, R.C., Kobayashi, Y., Neimanis, S., Walker, D.A., Heber, U. (1982) Co-operative activation of chloroplast fructose-1,6-bisphosphatase by reductant, pH and substrate. Biochim. Biophys. Acta 682, 168–178Google Scholar
  13. Lilley, R. McC., Walker, D.A. (1974) The reduction of 3-phosphogylcerate by reconstituted chloroplasts extracts. Biochim. Biophys. Acta 368, 269–278Google Scholar
  14. McGilvery, R.W. (1965) Fructose-1,6-diphosphate. Acidic dissociation constants, chelation with magnesium, and optical rotatory dispersion. Biochemistry 4, 1924–1930Google Scholar
  15. Meunier, J.C., Buc, J., Soulie, J.M., Piadel, J., Ricard, J. (1981) Substrate-binding isotherms of spinach chloroplastic fructose-1,6-bisphosphatase and the photoregulation of the Calvin cycle. Eur. J. Biochem. 113, 513–520Google Scholar
  16. Portis, A.R., Jr. (1981) Evidence of a low stromal Mg2+ concentration in intact chloroplasts in the dark. Plant Physiol. 67, 985–989Google Scholar
  17. Portis, A.R., Jr., Heldt, H.W. (1976) Light-dependent changes of the Mg2+ concentration in the stroma in relation to the Mg2+ dependency of CO2 fixation in intact chloroplasts. Biochim. Biophys. Acta 449, 434–466Google Scholar
  18. Portis, A.R., Jr., Chon, C.J., Mosbach, A., Heldt, H.W. (1977) Fructose- and sedoheptulosebisphosphatase. The sites of a possible control of CO2 fixation by light-dependent changes of the stromal Mg2+ concentration. Biochim. Biophys. Acta 461, 313–325Google Scholar
  19. Pradel, J., Soulie, J.M., Buc, J., Meunier, J.C., Ricard, J. (1981) On the activation of fructose-1,6-bisphosphatase of spinach chloroplasts and the regulation of the Calvin Cycle. Eur. J. Biochem. 113, 507–511Google Scholar
  20. Purczeld, P., Chon, C.J., Portis, A.R., Jr., Heldt, H.W., Heber, U. (1978) 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, 488–498Google Scholar
  21. Schürmann, P., Kobayashi, Y. (1983) Regulation of chloroplast fructose-1,6-bisphosphatase by the ferredoxin/thioredoxin system. Proc. 6th Int. Congr. on Photosynthesis, vol 3, pp. 629–632, Sybesma, C. ed. Nijhoff-Junk, The HagueGoogle Scholar
  22. Stitt, M., Mieskes, G., Söling, H.D., Heldt, H.W. (1982) On a possible role of fructose-2,6-bisphosphate in regulating photosynthetic metabolism in leaves. FEBS Lett. 145, 217–222Google Scholar
  23. Werdan, K., Heldt, H.W., Milovancev, M. (1975) The role of pH in the regulation of carbon fixation in the chloroplast stroma. Biochim. Biophys. Acta 396, 276–292Google Scholar
  24. Wirtz, W., Stitt, M., Heldt, H.W. (1982) Light activation of Calvin cycle enzymes as measured in pea leaves. FEBS Lett. 142, 223–226Google Scholar
  25. Woodrow, J.E., Murphy, D.J., Latzko, E. (1984) Regulation of stromal sedoheptulose 1,7-bisphosphatase activity by pH and Mg2+ concentration. J. Biol. Chem. 259, 3791–3795Google Scholar
  26. Zimmermann, G., Kelly, J., Latzko, E. (1976) Efficient purification and molecular properties of spinach fructose-1,6-bisphosphatase. Eur. J. Biochem. 70, 361–367Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • A. Gardemann
    • 1
  • D. Schimkat
    • 1
  • H. W. Heldt
    • 1
  1. 1.Institut für Biochemie der Pflanze der Universität GöttingenGöttingenFederal Republic of Germany

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