Plant Molecular Biology

, Volume 40, Issue 5, pp 825–834

Molecular cloning, expression, and functional characterization of a 2Cys-peroxiredoxin in Chinese cabbage

  • Na Eun Cheong
  • Yeon Ok Choi
  • Kyun Oh Lee
  • Woe Yeon Kim
  • Bae Gyo Jung
  • Yong Hun Chi
  • Jin Sook Jeong
  • Kanghwa Kim
  • Moo Je Cho
  • Sang Yeol Lee
Article

DOI: 10.1023/A:1006271823973

Cite this article as:
Cheong, N.E., Choi, Y.O., Lee, K.O. et al. Plant Mol Biol (1999) 40: 825. doi:10.1023/A:1006271823973

Abstract

A cDNA (C2C-Prx) corresponding to a 2Cys-peroxiredoxin (2Cys-Prx) was isolated from a leaf cDNA library of Chinese cabbage. The predicted amino acid sequence of C2C-Prx has 2 conserved cysteines and several peptide domains present in most of the 2Cys-Prx subfamily members. It shows the highest sequence homology to the 2Cys-Prx enzymes of spinach (88%) and Arabidopsis (86%). Southern analysis using the cDNA insert of C2C-Prx revealed that it consists of a small multigene family in Chinese cabbage genome. RNA blot analysis showed that the gene was predominantly expressed in the leaf tissue of Chinese cabbage seedlings, but the mRNA was generally expressed in most tissues of mature plant, except roots. The expression of C2C-Prx was slightly induced by treatment with H2O2 (100μM) or Fe3+/O2/DTT oxidation system, but not by ABA (50 μM) or GA3 (10 μM). The C2C-Prx is encoded as a preprotein of 273 amino acids containing a putative chloroplast-targeting signal of 65 amino acids at its N-terminus. The N-terminally truncated recombinant protein (ΔC2C-Prx) migrates as a dimer in a non-reducing SDS-polyacrylamide gel and as a monomer in a reducing condition. The ΔC2C-Prx shows no immuno cross-reactivity to antiserum of the yeast thiol-specific antioxidant protein, and vice versa. The ΔC2C-Prx prevents the inactivation of glutamine synthetase and the DNA cleavage in the metal-catalyzed oxidation system. In the yeast thioredoxin system containing thioredoxin reductase, thioredoxin, and NADPH, the ΔC2C-Prx exhibits peroxidase activity on H2O2.

2Cys-peroxiredoxinChinese cabbageexpressionfunctional characterizationgene cloning

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Na Eun Cheong
    • 1
  • Yeon Ok Choi
    • 2
  • Kyun Oh Lee
    • 1
  • Woe Yeon Kim
    • 1
  • Bae Gyo Jung
    • 1
  • Yong Hun Chi
    • 1
  • Jin Sook Jeong
    • 3
  • Kanghwa Kim
    • 3
  • Moo Je Cho
    • 1
  • Sang Yeol Lee
    • 1
  1. 1.Department of BiochemistryCollege of Natural SciencesKorea
  2. 2.Plant Molecular Biology and Biotechnology Research CenterGyeongsang National UniversityChinjuKorea
  3. 3.Department of Food and NutritionChonnam National UniversityKwangjuKorea