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Sucrose-phosphate synthase is regulated via metabolites and protein phosphorylation in potato tubers, in a manner analogous to the enzyme in leaves

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Abstract

(i) Sucrose-phosphate synthase (SPS) was purified 40-fold from stored potato (Solanum tuberosum L.) tubers to a final specific activity of 33–70 nkat·(mg protein)−1 via batch elution from diethylaminoethyl (DEAE)-sephacel, polyethylene glycol (PEG) precipitation and Mono Q anion-exchange chromatography. (ii) Immunoblotting revealed a major and a minor band with molecular weights of 124.8 kDa and 133.5 kDa, respectively. Both bands were also present in extracts prepared in boiling SDS to exclude proteolysis. No smaller polypeptides were seen, except when the preparations were incubated before application on a polyacrylamide gel. (iii) The enzyme preparation was activated by glucose-6-phosphate and inhibited by inorganic phosphate. Both effectors had a large effect on the K m (fructose-6-phosphate) and the K m (uridine-5-diphosphoglucose) with phosphate acting antagonistically to glucose-6-phosphate. (iv) Preincubation of potato slices with low concentrations of okadaic acid or microcystin resulted in a three- to fourfold decrease in the activity of SPS when the tissue was subsequently extracted and assayed. The decrease was especially marked when the assay contained low concentrations of substrates and glucose-6-phosphate, and inorganic phosphate was included. Preincubation with mannose or in high osmoticum resulted in an increase of SPS activity. (v) Analogous changes were observed in germinating Ricinus communis L. seedlings. After preincubation of the cotyledons in glucose, high SPS activity could be measured, whereas okadaic acid, omission of glucose, or addition of phosphate or sucrose led to a large decrease of SPS activity in the “selective” assay. (vi) It is argued that SPS from non-photosynthetic tissues is regulated by metabolites and by protein phosphorylation in an analogous manner to the leaf enzyme.

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Abbreviations

Fru6P:

fructose-6-phosphate

Glc6P:

glucose-6-phosphate

Pi:

inorganic phosphate

PGI:

phosphoglucose isomerase

PP2A:

phosphoprotein phosphatase 2A

PEG:

polyethyleneglycol

SPS:

sucrose-phosphate synthase

UDPGlc:

uridine-5-diphosphoglucose

References

  • Bruneau, J.-M., Worrell, A.C., Cambou, B., Lando, D., Voelker, T.A. (1991) Sucrose phosphate synthase, a key enzyme for sucrose biosynthesis in plants. Plant Physiol. 96, 473–478

    Google Scholar 

  • Cardini, C.E., Slabnik, E., Frydman, R.B. (1968) Some properties of potato tuber UDP glucose: d-fructose-2-glucosyltransferase and UDP glucose: d-fructose-6-phosphate-2-glucosyltransferase. Plant Physiol. 43, 1063–1068

    Google Scholar 

  • Dancer, J.E., Matzfeld, W.D., Stitt, M. (1990) Cytosolic cycles regulate the turnover of sucrose in heterotrophic cell-suspension cultures of Chenopodium rubrum L. Planta 182, 223–251

    Google Scholar 

  • Doehlert, D.C., Huber, S.C. (1983) Regulation of spinach leaf sucrose-phosphate synthase by glucose-6-phosphate, inorganic phosphate and pH. Plant Physiol. 73, 989–994

    Google Scholar 

  • Doehlert, D.C., Huber, S.C. (1985) The role of sulfhydryl groups in the regulation of spinach leaf sucrose-phosphate synthase. Biochim. Biophys. Acta 830, 276–273

    Google Scholar 

  • Geigenberger, P., Stitt, M. (1991) A futile cycle of sucrose synthase and degradation is involved in regulating partitioning between sucrose, starch and respiration in cotyledons of germinating Ricinus communis L. seedlings when phloem transport is inhibited. Planta 185, 81–90

    CAS  Google Scholar 

  • Geigenberger, P., Stitt, M. (1993) Sucrose synthase catalyses a readily reversible reaction in vivo in developing potato tubers and other plant tissues. Planta 189, 329–339

    CAS  Google Scholar 

  • Guy, C.L. (1990) Cold acclimation and freezing stress tolerance. Annu. Rev. Plant Physiol. Mol. Biol. 41, 187–223

    Google Scholar 

  • Hajirezeai, M., Stitt, M. (1991) Contrasting roles for pyrophosphate: fructose-6-phosphate phosphotransferase during aging of tissue slices from potato tubers and carrot slices. Plant Sci. 77, 177–183

    Google Scholar 

  • Hawker, J.S. (1985) Sucrose. In: The biochemistry of storage carbohydrates in green plants, pp. 1–51, Dey, P.M., Dixon, R.A. eds. Academic Press, New York

    Google Scholar 

  • Hubbard, N.L., Pharr, M.D., Huber S.C. (1990) Role of sucrose phosphate synthase in ripening bananas and its relation to the respiratory climacteric. Plant Physiol. 94, 201–208

    Google Scholar 

  • Huber, S.C., Huber, J.L.A. (1991) Regulation of maize leaf sucrose-phosphate synthase by protein phosphorylation. Plant Cell Physiol. 32, 319–326

    Google Scholar 

  • Huber, S.C., Huber, J.L.A., Nielsen, T.H. (1989) Protein phosphorylation as a mechanism for regulation of spinach leaf sucrose-phosphate synthase activity. Arch. Biochem. Biophys. 270, 681–690

    Google Scholar 

  • Jelitto, T., Sonnewald, U., Willmitzer, L., Hajirezeai, M., Stitt, M. (1992) Inorganic pyrophosphate content and metabolites in potato and tobacco plants expressing E. coli pyrophosphatase in their cytosol. Planta 188, 238–244

    Google Scholar 

  • Kalt-Torres, W., Kerr, P.S., Huber, S.C. (1987) Isolation and characterization of multiple forms of maize leaf sucrose-phosphate synthase. Plant Physiol. 70, 653–658

    Google Scholar 

  • Krüger, N. (1990) Carbohydrate synthesis and degradation. In: Plant physiology, biochemistry and molecular biology, pp. 59–76, Dennis, D.T., Turpin, D.M., eds. Lorgmar, Singapore

    Google Scholar 

  • MacRae, E., Quick, W.P., Benker, C., Stitt, M. (1992) Carbohydrate metabolism during ripening of kiwi fruit. Planta 188, 314–323

    Google Scholar 

  • Neuhaus, H.E., Quick, W.P., Siegl, G., Stitt, M. (1990) Control of photosynthate partitioning in spinach leaves. Planta 181, 583–592

    Google Scholar 

  • Nomura, T., Akazawa, T. (1974) Enzyme mechanism of starch breakdown in germinating rice seeds. V. Sucrose phosphate synthase in the scutellum. Plant Cell Physiol. 15, 477–483

    Google Scholar 

  • Plaxton, W.C., Preiss, J. (1987) Purification and properties of non-proteolytic degraded ADP glucose pyrophosphorylase from maize endosperm. Plant Physiol. 83, 105–112

    Google Scholar 

  • Quick, W.P., Siegl, G., Neuhaus, H.E., Stitt, M. (1989) Short-term water stress leads to a stimulation of sucrose synthesis by activating sucrose phosphate synthase. Planta 177, 535–546

    CAS  Google Scholar 

  • Salerno, G.L., Pontis, H.G. (1978) Sucrose phosphate synthase, separation from sucrose synthase and study of its properties. Planta 142, 41–48

    Google Scholar 

  • Salvucci, M.E., Drake, R.R., Haley, B.E. (1990) Purification and photoaffinity labelling of sucrose-phosphate synthase from spinach leaves. Arch. Biochem. Biophys. 281, 212–218

    Google Scholar 

  • Siegl, G., Mackintosh, C., Stitt, M. (1990) Sucrose phosphate synthase is dephosphorylated by protein phosphatase 2A in spinach leaves. FEBS Lett. 270, 198–202

    Google Scholar 

  • Siegl, G., Stitt, M. (1990) Partial purification of two forms of spinach leaf sucrose-phosphate synthase which differ in their kinetic properties. Plant Sci. 66, 205–210

    Google Scholar 

  • Sonnewald, U., Quick, W.P., MacRae, E., Krause, K.-P., Stitt, M. (1993) Purification, cloning and expression of spinach leaf sucrose-phosphate synthase in E. coli. Planta 189, 174–181

    Google Scholar 

  • Stitt, M., Huber, S.C., Kerr, P. (1987) Control of photosynthetic sucrose synthesis. In: The biochemistry of plants, vol.10, pp. 328–429, Hatch, M.D., Boardman, N.R. eds. Academic Press, New York

    Google Scholar 

  • Stitt, M., Wilke, I., Feil, R., Heldt, H.W. (1988) Coarse control of sucrose phosphate synthase in leaves: alterations of the kinetic properties in response to the rate of photosynthesis and accumulation of sucrose. Planta 174, 217–230

    Google Scholar 

  • Walker, J.L.A., Huber, S.C. (1989a) Purification and preliminary characterization of sucrose-phosphate synthase using monoclonal antibodies. Plant Physiol. 89, 518–524

    Google Scholar 

  • Walker, J.L.A., Huber, S.C. (1989b) Regulation of sucrose-phosphate synthase activity in spinach leaves by protein level and covalent modification. Planta 117, 116–120

    Google Scholar 

  • Weiner, H., McMichael, R.W., Huber, S.C. (1992) Identification of factors regulating the phosphorylation status of sucrose-phosphate synthase in vivo. Plant Physiol. 99, 1435–1442

    Google Scholar 

  • Zrenner, R., Stitt, M. (1991) Comparison of the effect of rapidly and gradually developing water-stress on carbohydrate metabolism in spinach leaves. Plant Cell Environ. 14, 939–946

    Google Scholar 

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Author notes

  1. Ralph Reimholz

    Present address: Botanisches Institut, Im Neuenheimer Feld 360, D-69120, Heidelberg, Germany

Authors and Affiliations

  1. Lehrstuhl für Pflanzenphysiologie, Universität Bayreuth, Universitätsstrasse 30, D-95447, Bayreuth, Germany

    Ralph Reimholz

  2. Botanisches Institut, Im Neuenheimer Feld 360, D-69120, Heidelberg, Germany

    Peter Geigenberger & Mark Stitt

Authors
  1. Ralph Reimholz
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  2. Peter Geigenberger
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  3. Mark Stitt
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Additional information

This work was supported by the Deutsche Forschungsgemeinschaft, the BMFT and Sandoz AG, Basel, Switzerland. We are grateful to Prof. E. Beck (Pflanzenphysiologie, Bayreuth, Germany) for providing us with laboratory facilities, and to Dr. U. Sonnewald (Institut für Genbiologische Forschung, Berlin, Germany) for many discussions and providing us with unpublished data.

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Reimholz, R., Geigenberger, P. & Stitt, M. Sucrose-phosphate synthase is regulated via metabolites and protein phosphorylation in potato tubers, in a manner analogous to the enzyme in leaves. Planta 192, 480–488 (1994). https://doi.org/10.1007/BF00203585

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