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A novel NADPH:diamide oxidoreductase activity in Arabidopsis thaliana P1 ζ-crystallin

(2000) EUROPEAN JOURNAL OF BIOCHEMISTRY. 267(12). p.3661-3671
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Abstract
The zeta-crystallin (ZCr) gene P1 of Arabidopsis thaliana, known to confer tolerance toward the oxidizing drug 1,1'-azobis(N,N-dimethylformamide) (diamide) to yeast [Babiychuk, E., Kushnir, S., Belles-Boix, E., Van Montagu, M. & Inze, D. (1995) J. Biol. Chem. 270, 26224], was expressed in Escherichia coli to characterize biochemical properties of the P1-zeta-crystallin (P1-ZCr). Recombinant P1-ZCr, a noncovalent dimer, showed NADPH:quinone oxidoreductase activity with specificity to quinones similar to that of guinea-pig ZCr. P1-ZCr also catalyzed the divalent reduction of diamide to 1,2-bis(N,N-dimethylcarbamoyl)hydrazine, with a k(cat) comparable with that for quinones. Two other azodicarbonyl compounds also served as substrates of P1-ZCr. Guinea-pig ZCr, however, did not catalyze the azodicarbonyl reduction. Hence, plant ZCr is distinct from mammalian ZCr, and can be referred to as NADPH:azodicarbonyl/quinone reductase. The quinone-reducing reaction was accompanied by radical chain reactions to produce superoxide radicals, while the azodicarbonyl-reducing reaction was not. Specificity to NADPH, as judged by k(cat)/K-m, was >1000-fold higher than that to NADH both for quinones and diamide. N-Ethylmaleimide and p-chloromercuribenzoic acid inhibited both quinone-reducing and diamide-reducing activities. Both NADPH and NADP(+) suppressed the inhibition, but NADH did not, suggesting that sulfhydryl groups reside in the binding site for the phosphate group on the adenosine moiety of NADPH. The diamide-reducing activity of P1-ZCr accounts for the tolerance of P1-overexpressing yeast to diamide. Other possible physiological functions of P1-ZCr in plants are discussed.
Keywords
diamide, Arabidopsis thaliana, P1 protein, quinone reductase, zeta-crystallin, GUINEA-PIG LENS, OXIDIZING DRUG DIAMIDE, FATTY-ACID SYNTHASE, QUINONE OXIDOREDUCTASE, ESCHERICHIA-COLI, OXIDATIVE STRESS, ALCOHOL DEHYDROGENASES, HYDROGEN-PEROXIDE, GENE-EXPRESSION, DT-DIAPHORASE

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Citation

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Chicago
Mano, Jun’ichi, Elena Babiychuk, Enric Belles-Boix, Jun Hiratake, Akira Kimura, Dirk Inzé, Sergei Kushnir, and Kozi Asada. 2000. “A Novel NADPH:diamide Oxidoreductase Activity in Arabidopsis Thaliana P1 Ζ-crystallin.” European Journal of Biochemistry 267 (12): 3661–3671.
APA
Mano, Jun’ichi, Babiychuk, E., Belles-Boix, E., Hiratake, J., Kimura, A., Inzé, D., Kushnir, S., et al. (2000). A novel NADPH:diamide oxidoreductase activity in Arabidopsis thaliana P1 ζ-crystallin. EUROPEAN JOURNAL OF BIOCHEMISTRY, 267(12), 3661–3671.
Vancouver
1.
Mano J, Babiychuk E, Belles-Boix E, Hiratake J, Kimura A, Inzé D, et al. A novel NADPH:diamide oxidoreductase activity in Arabidopsis thaliana P1 ζ-crystallin. EUROPEAN JOURNAL OF BIOCHEMISTRY. 2000;267(12):3661–71.
MLA
Mano, Jun’ichi, Elena Babiychuk, Enric Belles-Boix, et al. “A Novel NADPH:diamide Oxidoreductase Activity in Arabidopsis Thaliana P1 Ζ-crystallin.” EUROPEAN JOURNAL OF BIOCHEMISTRY 267.12 (2000): 3661–3671. Print.
@article{125000,
  abstract     = {The zeta-crystallin (ZCr) gene P1 of Arabidopsis thaliana, known to confer tolerance toward the oxidizing drug 1,1'-azobis(N,N-dimethylformamide) (diamide) to yeast [Babiychuk, E., Kushnir, S., Belles-Boix, E., Van Montagu, M. \& Inze, D. (1995) J. Biol. Chem. 270, 26224], was expressed in Escherichia coli to characterize biochemical properties of the P1-zeta-crystallin (P1-ZCr). Recombinant P1-ZCr, a noncovalent dimer, showed NADPH:quinone oxidoreductase activity with specificity to quinones similar to that of guinea-pig ZCr. P1-ZCr also catalyzed the divalent reduction of diamide to 1,2-bis(N,N-dimethylcarbamoyl)hydrazine, with a k(cat) comparable with that for quinones. Two other azodicarbonyl compounds also served as substrates of P1-ZCr. Guinea-pig ZCr, however, did not catalyze the azodicarbonyl reduction. Hence, plant ZCr is distinct from mammalian ZCr, and can be referred to as NADPH:azodicarbonyl/quinone reductase. The quinone-reducing reaction was accompanied by radical chain reactions to produce superoxide radicals, while the azodicarbonyl-reducing reaction was not. Specificity to NADPH, as judged by k(cat)/K-m, was {\textrangle}1000-fold higher than that to NADH both for quinones and diamide. N-Ethylmaleimide and p-chloromercuribenzoic acid inhibited both quinone-reducing and diamide-reducing activities. Both NADPH and NADP(+) suppressed the inhibition, but NADH did not, suggesting that sulfhydryl groups reside in the binding site for the phosphate group on the adenosine moiety of NADPH. The diamide-reducing activity of P1-ZCr accounts for the tolerance of P1-overexpressing yeast to diamide. Other possible physiological functions of P1-ZCr in plants are discussed.},
  author       = {Mano, Jun'ichi and Babiychuk, Elena and Belles-Boix, Enric and Hiratake, Jun and Kimura, Akira and Inz{\'e}, Dirk and Kushnir, Sergei and Asada, Kozi},
  issn         = {0014-2956},
  journal      = {EUROPEAN JOURNAL OF BIOCHEMISTRY},
  keyword      = {diamide,Arabidopsis thaliana,P1 protein,quinone reductase,zeta-crystallin,GUINEA-PIG LENS,OXIDIZING DRUG DIAMIDE,FATTY-ACID SYNTHASE,QUINONE OXIDOREDUCTASE,ESCHERICHIA-COLI,OXIDATIVE STRESS,ALCOHOL DEHYDROGENASES,HYDROGEN-PEROXIDE,GENE-EXPRESSION,DT-DIAPHORASE},
  language     = {eng},
  number       = {12},
  pages        = {3661--3671},
  title        = {A novel NADPH:diamide oxidoreductase activity in Arabidopsis thaliana P1 \ensuremath{\zeta}-crystallin},
  url          = {http://dx.doi.org/10.1046/j.1432-1327.2000.01398.x},
  volume       = {267},
  year         = {2000},
}

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