Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The ferredoxin-thioredoxin system of a green alga, Chlamydomonas reinhardtii

Identification and characterization of thioredoxins and ferredoxin-thioredoxin reductase components

  • 66 Accesses

  • 1 Citations

Abstract

The components of the ferredoxin-thioredoxin (FT) system of Chlamydomonas reinhardtii have been purified and characterized. The system resembled that of higher plants in consisting of a ferredoxin-thioredoxin reductase (FTR) and two types of thioredoxin, a single f and two m species, m1 and m2. The Chlamydomonas m and f thioredoxins were antigenically similar to their higher-plant counterparts, but not to one another. The m thioredoxins were recognized by antibodies to both higher-plant m and bacterial thioredoxins, whereas the thioredoxin f was not. Chlamydomonas thioredoxin f reacted, although weakly, with the antibody to spinach thioredoxin f. The algal thioredoxin f differed from thioredoxins studied previously in behaving as a basic protein on ion-exchange columns. Purification revealed that the algal thioredoxins had molecular masses (Mrs) typical of thioredoxins from other sources, m1 and m2 being 10700 and f 11 500. Chlamydomonas FTR had two dissimilar subunits, a feature common to all FTRs studied thus far. One, the 13-kDa (“similar”) subunit, resembled its counterpart from other sources in both size and antigenicity. The other, 10-kDa (“variable”) sub-unit was not recognized by antibodies to any FTR tested. When combined with spinach, (Spinacia oleracea L.) thylakoid membranes, the components of the FT system functioned in the light activation of the standard target enzymes from chloroplasts, corn (Zea mays L.) NADP-malate dehydrogenase (EC 1.1.1.82) and spinach fructose 1,6-bisphosphatase (EC 3.1.3.11) as well as the chloroplast-type fructose 1,6-bisphosphatase from Chlamydomonas. Activity was greatest if ferredoxin and other components of the FT system were from Chlamydomonas. The capacity of the Chlamydomonas FT system to activate autologous FBPase indicates that light regulates the photosynthetic carbon metabolism of green algae as in other oxygenic photosynthetic organisms.

This is a preview of subscription content, log in to check access.

Abbreviations

DEAE:

diethylaminoethyl

ELISA:

enzyme-linked immunosorption assay

FBPase:

fructose 1,6-bisphosphatase

Fd:

ferredoxin

FPLC:

fast protein liquid chromatography

FTR:

ferredoxin-thioredoxin reductase

FT system:

ferredoxin-thioredoxin system

kDa:

kilodaltons

Mr :

relative molecular mass

NADP-MDH:

NADP-malate dehydrogenase

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

References

  1. Anderson, L.E., Avron, M. (1976) Light modulation of enzyme activity in chloroplasts. Plant Physiol. 57, 209–213

  2. Bassham, J.A. (1979) The reductive pentose phosphate cycle and its regulation. In: Encyclopedia of plant physiology, N.S., vol. 6. Photosynthesis II, Photosynthesis carbon metabolism and related processes, pp. 9–30, Gibbs, E., Latzko, E., eds. Springer, Berlin Heidelberg New York

  3. Buc, J., Rivière, M., Gontero, B., Sauve, P., Meunier, J.-C., Ricard, J. (1984) Affinity chromatography, on fructose-bisphosphatase-Sepharose, of two chloroplastic thioredoxins F. Eur. J. Biochem. 140, 199–202

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

  5. Buchanan, B.B., Hartman H., Syvanen, M. (1989) Evolutionary history of chloroplast thioredoxins f and m. (Abstr.) Plant Physiol. 98, Suppl., 174

  6. Cossar, J.D., Rowell, P., Stewart, W.D.P. (1984) Thioredoxin as a modulator of glucose-6-phosphate dehydrogenase in a N2-fixing cyanobacterium. J. Gen. Microbiol. 130, 991–998

  7. 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, 1–15

  8. Crawford, N.A., Droux, M., Kosower, N.S., Buchanan, B.B. (1989) Evidence for the function of ferredoxin/thioredoxin system in the reductive activation of target enzymes of isolated intact chloroplasts. Arch. Biochem. Biophys. 271, 223–239

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

  10. Decottignies, P., Schmitter, J., Miginiac-Maslow, M., Le Marechal, P., Jacquot, J.-P., Gadal, P. (1988) Primary structure of the light dependent regulatory site of corn NADP-malate dehydrogenase. J. Biol. Chem. 263, 11780–11785

  11. Droux, M., Crawford, N.A., Buchanan, B.B. (1987a) Mechanism of thioredoxin-linked activation of chloroplast fructose 1,6-bisphosphatase. C.R. Acad. Sci. Paris 305, Sér. III, 335–341

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

  13. Droux, M., Miginiac-Maslow, M., Jacquot, J.P., Gadal, P., Crawford, N.A., Kosower, N.S., Buchanan, B.B. (1987c) Ferredoxin-thioredoxin reductase: a catalytically active dithiol group links photoreduced ferredoxin to thioredoxin functional in photosynthetic enzyme regulation. Arch. Biochem. Biophys. 256, 372–380

  14. Florencio, F.J., Yee, B.C., Johnson, T.C., Buchanan, B.B. (1988) An NADP/thioredoxin system in leaves: purification and characterization of NADP-thioredoxin reductase and thioredoxin h from spinach. Arch. Biochem. Biophys. 266, 496–507

  15. Ford, D.M., Jablonski, P.P., Mohamed, A.H., Anderson, L.E. (1987) Protein modulase appears to be a complex of ferredoxin, ferredoxin/thioredoxin reductase, and thioredoxin. Plant Physiol. 83, 628–632

  16. Hashimoto, F., Horigome, T., Kanbayashi, M., Yoshida, K., Sugano, H. (1983) An improved method for separation of low molecular weight polypeptides in sodium dodecyl sulfate-poly-acrylamide gel. Anal. Biochem. 129, 192–199

  17. Hirasawa, M., Droux, M., Gray, K.A., Boyer, J.M., Davis, D.J., Buchanan, B.B., Knaff, D.B. (1988) Ferredoxin-thioredoxin reductase: properties of its complex with ferredoxin. Biochim. Biophys. Acta 925, 1–8

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

  19. Huppe, H.C., Buchanan, B.B. (1989) Activation of a chloroplast-type of fructose bisphosphatase from Chlamydomonas reinhardtii by light-mediated agents. Z. Naturforsch. 44c, 487–494

  20. Hutcheson, S.W., Buchanan, B.B. (1983) Enzyme regulation in crassulacean acid metabolism photosynthesis: studies on the ferredoxin/thioredoxin system of Kalanchoë daigremontiana. Plant Physiol. 72, 870–876

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

  22. Jacquot, J.-P., Vidal, J., Gadal, P., Schürmann, P. (1978) Evidence for the existence of several enzyme specific thioredoxins in plants. FEBS Lett. 96, 243–246

  23. 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

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

  25. Kamo, M., Tsugita, A., Weissner, C., Wedel, N., Bartling, D., Hermenn, R.G., Aguilar, F., Garrdet-Salvi, L., Schürmann, P. (1989) Primary structure of spinach chloroplasts thioredoxin f: protein sequencing and analysis of complete cDNA clones for spinach chloroplast thioredoxin f. Eur. J. Biochem. 182, 315–322

  26. Langlotz, P., Wagner, W., Follmann, H. (1986a) A large chloroplast thioredoxin f found in green algae. Z. Naturforsch. 41c, 275–283

  27. Langlotz, P., Wagner, W., Follmann H. (1986b) Green algae (Scenedesmus obliquus) contain three thioredoxins of regular size. Z. Naturforsch. 41c, 979–987

  28. 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

  29. Marcus, F., Moberly, L., Latshaw, S.P. (1988) Comparative amino acid sequence of fructose 1,6-bisphosphatase: Identification of a region unique to the light-regulated chloroplast enzyme. Proc. Natl. Acad. Sci. USA 85, 5379–5363

  30. Miki, J., Maeda, M., Mukohata, Y., Futai, M. (1988) The γ-subunit of ATP synthase from spinach chloroplasts. Primary structure from the cloned sequence. FEBS Lett. 232, 221–226

  31. 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. 707–713, Edelman, M., Hallick, R.B., Chua, N-H., eds. Elsevier, New York

  32. Pharmacia Fine Chemicals (1979) Affinity chromatography: Principles and methods. Ljungforetagen A.B., Örebro, Sweden

  33. Pla, A., Lopez-Gorge, J. (1981) Thioredoxin/fructose-1,6-bisphos-phatase affinity in the enzyme activation by the ferredoxin-thioredoxin system. Biochim. Biophys. Acta 636, 113–118

  34. Porter, M.A., Stringer, C.D., Hartman, F.C. (1988) Characterization of the regulatory site of phosphoribulokinase. J. Biol. Chem. 262, 123–129

  35. Raines, C.A., Lloyd, J.C., Longstaff, M., Bradley, D., Dyer, T. (1988) Chloroplast fructose 1,6-bisphosphatase: the product of a mosaic gene. Nucleic Acids Res. 16, 7931–7942

  36. Scheibe, R. (1987) NADP+-malate dehydrogenase in C3 plants: regulation and role of a light-activated enzyme. Physiol. Plant. 71, 393–400

  37. Schmitter, J.-M., Jacquot, J.-P., de Lamotte-Guéry, F., Beauvallet, C., Dutka, S., Gadal, P., Decottignies, P. (1988) Purification, properties and complete amino acid sequence of the ferredoxin from a green alga, Chlamydomonas reinhardtii. Eur. J. Biochem. 172, 405–412

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

  39. Tsugita, A., Maeda, K., Schürmann, P. (1983) Spinach chloroplast thioredoxins in evolutionary drift. Biochem. Biophys. Res. Commun. 115, 1–7

  40. Wagner A., Follmann, H. (1977) A thioredoxin from green algae. Biochem. Biophys. Res. Commun. 77, 1044–1050

  41. Whittaker, M.W., Gleason (1984) Isolation and characterization of thioredoxin f from the filamentous cyanobacteria, Anabaena sp. 7119. J. Biol. Chem. 259, 14088–14093

  42. Wolosiuk, R.A., Crawford, N.A., Yee, B.C., Buchanan, B.B. (1979) Isolation of three thioredoxins from spinach leaves. J. Biol. Chem. 254, 1627–1632

  43. 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

Download references

Author information

Correspondence to Bob B. Buchanan.

Additional information

This work was supported in part by a grant from the National Aeronautics and Space Administration. We would like to thank Don Carlson and Jacqueline Girard for their assistance with cell cultures.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Huppe, H.C., de Lamotte-Guéry, F., Jacquot, J. et al. The ferredoxin-thioredoxin system of a green alga, Chlamydomonas reinhardtii . Planta 180, 341–351 (1990). https://doi.org/10.1007/BF00198785

Download citation

Key words

  • Chlamydomonas
  • Chlorophyta
  • Ferredoxin-thioredoxin reductase
  • Light and thioredoxin
  • Thioredoxin