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Thioredoxin and NADP-thioredoxin reductase from cultured carrot cells


Dark-grown carrot (Daucus carota L.) tissue cultures were found to contain both protein components of the NADP/thioredoxin system—NADP—thioredoxin reductase and the thioredoxin characteristic of heterotrophic systems, thioredoxin h. Thioredoxin h was purified to apparent homogeneity and, like typical bacterial counterparts, was a 12-kdalton (kDa) acidic protein capable of activating chloroplast NADP-malate dehydrogenase (EC more effectively than fructose-1,6-bisphosphatase (EC NADP-thioredoxin reductase (EC was partially purified and found to be an arsenite-sensitive enzyme composed of two 34-kDa subunits. Carrot NADP-thioredoxin reductase resembled more closely its counterpart from bacteria rather than animal cells in acceptor (thioredoxin) specificity. Upon greening of the cells, the content of NADP-thioredoxin-reductase activity, and, to a lesser extent, thioredoxin h decreased. The results confirm the presence of a heterotrophic-type thioredoxin system in plant cells and raise the question of its physiological function.

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dithiolbis(2-nitrobenzoic acid)




terredoxin-thioredoxin, reductase


NADP-malate dehydrogenase


NADP-thioredoxin reductase


sodium-dodecyl sulfate


  1. Anderson, B.L., Berry, R.W., Telser, A. (1983) A sodium dodecyl sulfate-polyacrylamide gel electrophoresis system that separates peptides and proteins in the molecular weight range of 2,500 to 90,000. Anal. Biochem. 132, 365–375

  2. Appelquist, L.A., Stumpf, P.K., von Wettstein, D. (1968) Lipid synthesis and ultrastructure of isolated barley chloroplasts. Plant Physiol. 43, 163–187

  3. Berstermann, A., Vogt, K., Follmann, H. (1983) Plant seeds contain several thioredoxins of regular size. Eur. J. Biochem. 131, 339–344

  4. Bradford, M.M. (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principal of protein dye binding. Anal. Biochem. 72, 248–254

  5. Buchanan, B.B. (1980) Role of light in chloroplast enzymes. Annu. Rev. Plant Physiol. 31, 341–374

  6. Buchanan, B.B. (1984) The ferredoxin/thioredoxin system: A key element in regulatory function of light in photosynthesis. BioScience 34, 378–383

  7. Clement-Metral, J.D., Höög, J.-O., Holmgreen, A. (1986) Characterization of the thioredoxin system in a facultative phototroph, Rhodobacter sphaeroides Y. Eur. J. Biochem. 161, 119–126

  8. Crawford, N.A., Sutton, C.W., Yee, B.C., Johnson, T.C., Carlson, D.C., Buchanan, B.B. (1984) Contrasting modes of photosynthetic enzyme regulation in oxygenic and anoxygenic prokaryotes. Arch. Microbiol. 139, 124–129

  9. Crawford, N.A., Yee, B.C., Hutcheson, S.W., Wolosiuk, R.A., Buchanan, B.B. (1986) Enzyme regulation in C4 photosynthesis: purification, properties, and activities of thioredoxins from C4 and C3 plants. Arch. Biochem. Biophys. 244, 1–5

  10. Cséke, C., Buchanan, B.B. (1986) Regulation of the formation and utilization of photosynthate in leaves. Biochim. Biophys. Acta 853, 43–63

  11. Droux, M., Jacquot, J.P., Miginiac-Maslow, M., Gadal, P., Huet, J.C., Crawford, N.A., Yee, B.C., Buchanan, B.B. (1987) Ferredoxin-thioredoxin reductase: an iron-sulfur enzyme linking light to enzyme regulation in oxygenic photosynthesis. Purification and properties of the enzyme from C3, C4, and cyanobacteria species. Arch. Biochem. Biophys. 252, 426–439

  12. Gleason, F.K. (1986) Thioredoxin in cyanobacteria: structural and functional homology to thioredoxins from other organism. In: Thioredoxin and glutaredoxin systems: Structure and function, pp. 21–30, Holmgren, A., Jörnrall, H., Brändén, C.-I., Sjöberg, B.-M., eds. Raven Press, New York

  13. Hammel, K.E., Buchanan, B.B. (1981) Thioredoxin and ferredoxin-thioredoxin reductase activity occurring in a fermentative bacterium. FEBS Lett. 130, 88–92

  14. Hammel, K.E., Cornwell, K.E., Buchanan, B.B. (1983) Ferredoxin/flavoprotein-linked pathway for the reduction of thioredoxin. Proc. Natl. Acad. Sci. USA 80, 3681–3685

  15. Holmgren, A. (1977) Bovine thioredoxin system. J. Biol. Chem. 252, 4600–4606

  16. Holmgren, A. (1981) Pyridine nucleotide-disulfide oxidoreductases. In: Dehydrogenases requiring nicotinamide coenzymes, p. 149–180, Jeffrey, J., ed. Birkhäuser, Basel

  17. Holmgren, A. (1985) Thioredoxin. Annu. Rev. Biochem. 54, 237–271

  18. Huppe, H., Buchanan, B.B., Christophe de Lamotte-Guery, F., Jacquot, J.P. (1987) The ferredoxin/thioredoxin system from green alga, Chlamydomonas reinhardtii. In: Progress in photosynthesis research, pp. 443–446, Biggins, J., ed. Martinus Nijhoff, Boston

  19. Jacquot, J.-P. (1984) Post-translational modification of proteins in higher plant chloroplasts: enzyme regulation by thioldisulfide interchange. Physiol. Vég. 22, 487–507

  20. Jacquot, J.-P., Buchanan, B.B., Martin, F., Vidal, J. (1981) Enzyme regulation in C4 photosynthesis. Purification and properties of thioredoxin-linked NADP-malate dehydrogenase from corn leaves. Plant Physiol. 68, 300–304

  21. Jacquot, J.-P., Maudinas, B.G., Gadal, P. (1979) Occurrence of thioredoxin activity in the photosynthetic bacterium Rhodopseudomonas capsulata. Biochem. Biophys. Res. Commun. 90, 1371–1376

  22. Johnson, T.C., Crawford, N.A., Buchanan, B.B. (1984) Thioredoxin system of the photosynthetic anaerobe Chromatium vinosum. J. Bacteriol. 158, 1061–1069

  23. Laurent, T.C., Moore, E.D., Reichard, P. (1964) Enzymatic synthesis of deoxyribonucleotides. J. Biol. Chem. 239, 3436–3444

  24. Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685

  25. Maeda, K., Tsugita, A., Dalzoppo, D., Vilbois, F., Schürmann, P. (1986) Further characterization and amino acid sequence of m-type thioredoxins from spinach chloroplasts. Eur. J. Biochem. 154, 197–203

  26. Mathews, W.R., Johnson, R.S., Cornwell, K.L., Johnson, T.C., Buchanan, B.B., Biemann, K. (1987) Mass spectrometrically derived amino acid sequence of thioredoxin from Chlorobium, an evolutionarily prominent photosynthetic bacterium. J. Biol. Chem., in press

  27. Meister, A., Anderson, M.E. (1983) Glutathione. Annu. Rev. Biochem. 52, 711–760

  28. Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol. Plant. 15, 473–497

  29. Nishizawa, A.N., Yee, B.C., Buchanan, B.B. (1982) Chloroplast fructose-1,6-bisphosphatase from spinach leaves. In: Methods in chloroplast molecular biology, pp. 707–714, Edelman, M., Hallick, R.B., Chua, N.-H., eds. Elsevier Biochemical Press, New York

  30. Pigiet, V.P., Conley, R.R. (1977) Purification of thioredoxin reductase and glutathione reductase by affinity chromatography. J. Biol. Chem. 252, 6367–6372

  31. Schürmann, P. (1981) The ferredoxin thioredoxin system of chloroplasts: purification and characterization of its components. In: Proc. V Int. Congr. in Photosynthesis, vol 4., p. 273–280, Akoyunoglou, G., ed. Balaban International Science Service, Philadelphia

  32. Schürmann, P., Maeda, K., Tsugita, A. (1981) Isomers in thioredoxins of spinach chloroplasts. Eur. J. Biochem. 116, 37–45

  33. Spurr, A.R., Harris, W.M. (1968) Ultrastructure of chloroplasts and chromoplasts in Capsicum annuum. 1. Thylakoid membrane changes during fruit ripening. Am. J. Bot. 55, 1210–1223

  34. Spurr, A.R. (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J. Ultrastruct. Res. 26, 31–43

  35. Suske, G., Wagner, W., Follman, H. (1979) NADP-dependent thioredoxin reductase and a new thioredoxin from wheat. Z. Naturforsch. 34c, 214–221

  36. Tillman, P., Norman, C. (1983) Droperidol-fentanyl as an aid to blood collection in rabbits. Lab. Anim. Sci. 3, 181–182

  37. Tsang, M.L. (1981) Thioredoxin/glutaredoxin system of Chlorella. Plant Physiol. 68, 1098–1104

  38. Tsugita, A., Maeda, K., Schürmann, P. (1983) Spinach chloroplast thioredoxin in evolutionary drift. Biochem. Biophys. Res. Commun. 155, 1–7

  39. Wolosiuk, R.A., Schürmann, P., Buchanan, B.B. (1980) Thioredoxin and ferredoxin-thioredoxin reductase of spinach chloroplasts. Methods Enzymol. 69, 382–391

  40. Yee, B.C., de la Torre, A., Crawford, N.A., Lara, C., Carlson, D.E., Buchanan, B.B. (1981) The ferredoxin/thioredoxin system of enzyme regulation in a cyanobacterium. Arch. Microbiol. 130, 14–18

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Johnson, T.C., Cao, R.Q., Kung, J.E. et al. Thioredoxin and NADP-thioredoxin reductase from cultured carrot cells. Planta 171, 321–331 (1987).

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Key words

  • Cell culture (thioredoxin)
  • Daucus (thioredoxin)
  • NADP-thioredoxin reductase
  • Thioredoxin