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Thioredoxin and NADP-thioredoxin reductase from cultured carrot cells

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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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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Key words

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