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