Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1995 Jun;108(2):609–614. doi: 10.1104/pp.108.2.609

Enzymatic Evidence for a Complete Oxidative Pentose Phosphate Pathway in Chloroplasts and an Incomplete Pathway in the Cytosol of Spinach Leaves.

C Schnarrenberger 1, A Flechner 1, W Martin 1
PMCID: PMC157380  PMID: 12228497

Abstract

The intracellular localization of transaldolase, transketolase, ribose-5-phosphate isomerase, and ribulose-5-phosphate epimerase was reexamined in spinach (Spinacia oleracea L.) leaves. We found highly predominant if not exclusive localization of these enzyme activities in chloroplasts isolated by isopyknic centrifugation in sucrose gradients. Glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, glucose phosphate isomerase, and triose phosphate isomerase activity was present in the chloroplast fraction but showed additional activity in the cytosol (supernatant) fraction attributable to the cytosol-specific isoforms known to exist for these enzymes. Anion-exchange chromatography of proteins of crude extracts on diethylaminoethyl-Fractogel revealed only a single enzyme each for transaldolase, transketolase, ribose-5-phosphate isomerase, and ribulose-5-phosphate epimerase. The data indicate that chloroplasts of spinach leaf cells possess the complete complement of enzymes of the oxidative pentose phosphate path-way (OPPP), whereas the cytosol contains only the first two reactions, contrary to the widely held view that plants generally possess a cytosolic OPPP capable of cyclic function. The chloroplast enzymes transketolase, ribose-5-phosphate isomerase, and ribulose-5-phosphate epimerase appear to be amphibolic for the Calvin cycle and OPPP.

Full Text

The Full Text of this article is available as a PDF (642.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson L. E., Advani V. R. Chloroplast and cytoplasmic enzymes: three distinct isoenzymes associated with the reductive pentose phosphate cycle. Plant Physiol. 1970 May;45(5):583–585. doi: 10.1104/pp.45.5.583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BUTT V. S., BEEVERS H. The regulation of pathways of glucose catabolism in maize roots. Biochem J. 1961 Jul;80:21–27. doi: 10.1042/bj0800021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bassham J. A., Kirk M., Jensen R. G. Photosynthesis by isolated chloroplasts. I. Diffusion of labeled photosynthetic intermediates between isolated chloroplasts and suspending medium. Biochim Biophys Acta. 1968 Jan 15;153(1):211–218. doi: 10.1016/0005-2728(68)90162-x. [DOI] [PubMed] [Google Scholar]
  5. Bohlmann J., DeLuca V., Eilert U., Martin W. Purification and cDNA cloning of anthranilate synthase from Ruta graveolens: modes of expression and properties of native and recombinant enzymes. Plant J. 1995 Mar;7(3):491–501. doi: 10.1046/j.1365-313x.1995.7030491.x. [DOI] [PubMed] [Google Scholar]
  6. Dalling M. J., Tolbert N. E., Hageman R. H. Intracellular location of nitrate reductase and nitrite reductase. II. Wheat roots. Biochim Biophys Acta. 1972 Dec 14;283(3):513–519. doi: 10.1016/0005-2728(72)90267-8. [DOI] [PubMed] [Google Scholar]
  7. Graeve K., von Schaewen A., Scheibe R. Purification, characterization, and cDNA sequence of glucose-6-phosphate dehydrogenase from potato (Solanum tuberosum L.). Plant J. 1994 Mar;5(3):353–361. doi: 10.1111/j.1365-313x.1994.00353.x. [DOI] [PubMed] [Google Scholar]
  8. Görlach J., Beck A., Henstrand J. M., Handa A. K., Herrmann K. M., Schmid J., Amrhein N. Differential expression of tomato (Lycopersicon esculentum L.) genes encoding shikimate pathway isoenzymes. I. 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase. Plant Mol Biol. 1993 Nov;23(4):697–706. doi: 10.1007/BF00021525. [DOI] [PubMed] [Google Scholar]
  9. Heber U., Hallier U. W., Hudson M. A. Untersuchungen zur intrazellulären Verteilungen von Enzymen und Substraten in der Blattzelle. II. Lokalisation von Enzymen des reduktiven und dem oxydativen Pentosephosphat-Zyklus in den Chloroplasten und Permeabilität der Chloroplasten-Membran gegenüber Metaboliten. Z Naturforsch B. 1967 Nov;22(11):1200–1215. [PubMed] [Google Scholar]
  10. Herrmann K. M. The shikimate pathway as an entry to aromatic secondary metabolism. Plant Physiol. 1995 Jan;107(1):7–12. doi: 10.1104/pp.107.1.7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ireland R. J., Deluca V., Dennis D. T. Isoenzymes of pyruvate kinase in etioplasts and chloroplasts. Plant Physiol. 1979 May;63(5):903–907. doi: 10.1104/pp.63.5.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Krüger I., Schnarrenberger C. Purification, subunit structure and immunological comparison of fructose-bisphosphate aldolases from spinach and corn leaves. Eur J Biochem. 1983 Oct 17;136(1):101–106. doi: 10.1111/j.1432-1033.1983.tb07711.x. [DOI] [PubMed] [Google Scholar]
  13. Köpke-Secundo E., Molnar I., Schnarrenberger C. Isolation and characterization of the cytosolic and chloroplastic 3-phosphoglycerate kinase from spinach leaves. Plant Physiol. 1990 May;93(1):40–47. doi: 10.1104/pp.93.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Last R. L. The genetics of nitrogen assimilation and amino acid biosynthesis in flowering plants: progress and prospects. Int Rev Cytol. 1993;143:297–330. doi: 10.1016/s0074-7696(08)61878-1. [DOI] [PubMed] [Google Scholar]
  15. Latzko E., Zimmermann G., Feller U. Evidence for a hexosediphosphatase from the cytoplasm of spinach leaves. Hoppe Seylers Z Physiol Chem. 1974 Mar;355(3):321–326. doi: 10.1515/bchm2.1974.355.1.321. [DOI] [PubMed] [Google Scholar]
  16. Lebherz H. G., Leadbetter M. M., Bradshaw R. A. Isolation and characterization of the cytosolic and chloroplast forms of spinach leaf fructose diphosphate aldolase. J Biol Chem. 1984 Jan 25;259(2):1011–1017. [PubMed] [Google Scholar]
  17. McMorrow E. M., Bradbeer J. W. Separation, purification, and comparative properties of chloroplast and cytoplasmic phosphoglycerate kinase from barley leaves. Plant Physiol. 1990 Jun;93(2):374–383. doi: 10.1104/pp.93.2.374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Miernyk J. A., Dennis D. T. Isozymes of the glycolytic enzymes in endosperm from developing castor oil seeds. Plant Physiol. 1982 Apr;69(4):825–828. doi: 10.1104/pp.69.4.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miflin B. J., Beevers H. Isolation of intact plastids from a range of plant tissues. Plant Physiol. 1974 Jun;53(6):870–874. doi: 10.1104/pp.53.6.870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nishimura M., Beevers H. Isoenzymes of sugar phosphate metabolism in endosperm of germinating castor beans. Plant Physiol. 1981 Jun;67(6):1255–1258. doi: 10.1104/pp.67.6.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Niyogi K. K., Fink G. R. Two anthranilate synthase genes in Arabidopsis: defense-related regulation of the tryptophan pathway. Plant Cell. 1992 Jun;4(6):721–733. doi: 10.1105/tpc.4.6.721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Plaxton W. C. Molecular and immunological characterization of plastid and cytosolic pyruvate kinase isozymes from castor-oil-plant endosperm and leaf. Eur J Biochem. 1989 May 1;181(2):443–451. doi: 10.1111/j.1432-1033.1989.tb14745.x. [DOI] [PubMed] [Google Scholar]
  23. Rutner A. C. Spinach 5-phosphoribose isomerase. Purification and properties of the enzyme. Biochemistry. 1970 Jan 6;9(1):178–184. doi: 10.1021/bi00803a023. [DOI] [PubMed] [Google Scholar]
  24. Scheibe R., Geissler A., Fickenscher K. Chloroplast glucose-6-phosphate dehydrogenase: Km shift upon light modulation and reduction. Arch Biochem Biophys. 1989 Oct;274(1):290–297. doi: 10.1016/0003-9861(89)90441-4. [DOI] [PubMed] [Google Scholar]
  25. Schnarrenberger C., Krüger I. Distinction between Cytosol and Chloroplast Fructose-Bisphosphate Aldolases from Pea, Wheat, and Corn Leaves. Plant Physiol. 1986 Feb;80(2):301–304. doi: 10.1104/pp.80.2.301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schnarrenberger C., Oeser A., Tolbert N. E. Development of Microbodies in Sunflower Cotyledons and Castor Bean Endosperm during Germination. Plant Physiol. 1971 Nov;48(5):566–574. doi: 10.1104/pp.48.5.566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schnarrenberger C., Oeser A., Tolbert N. E. Two isoenzymes each of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in spinach leaves. Arch Biochem Biophys. 1973 Jan;154(1):438–448. doi: 10.1016/0003-9861(73)90077-5. [DOI] [PubMed] [Google Scholar]
  28. Schnarrenberger C., Oeser A. Two isoenzymes of glucosephosphate isomerase from spinach leaves and their intracellular compartmentation. Eur J Biochem. 1974 Jun 1;45(1):77–82. doi: 10.1111/j.1432-1033.1974.tb03531.x. [DOI] [PubMed] [Google Scholar]
  29. Simcox P. D., Dennis D. T. Isoenzymes of the Glycolytic and Pentose Phosphate Pathways in Proplastids from the Developing Endosperm of Ricinis communis L. Plant Physiol. 1978 Jun;61(6):871–877. doi: 10.1104/pp.61.6.871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. TABACHNICK M., SRERE P. A., COOPER J., RACKER E. The oxidative pentose phosphate cycle. III. The interconversion of ribose 5-phosphate, ribulose 5-phosphate and xylulose 5-phosphate. Arch Biochem Biophys. 1958 Apr;74(2):315–325. doi: 10.1016/0003-9861(58)90003-1. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES