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Thioredoxin and NADP-thioredoxin reductase from cultured carrot cells

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Abstract

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 1.1.1.82) more effectively than fructose-1,6-bisphosphatase (EC 3.1.3.11). NADP-thioredoxin reductase (EC 1.6.4.5) 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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Abbreviations

DTNB:

dithiolbis(2-nitrobenzoic acid)

FBPase:

fructose-1,6-bisphosphatase

FTR:

terredoxin-thioredoxin, reductase

NADP-MDH:

NADP-malate dehydrogenase

NTR:

NADP-thioredoxin reductase

SDS:

sodium-dodecyl sulfate

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Author notes

  1. Ri Qiang Cao

    Present address: Department of Biology, Nanjing University, Nanjing, China

Authors and Affiliations

  1. Division of Molecular Plant Biology, University of California, Hilgard Hall, 94720, Berkeley, CA, USA

    Thomas C. Johnson, Ri Qiang Cao, Jacob E. Kung & Bob B. Buchanan

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  1. Thomas C. Johnson
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  2. Ri Qiang Cao
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  3. Jacob E. Kung
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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). https://doi.org/10.1007/BF00398677

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  • DOI: https://doi.org/10.1007/BF00398677

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