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Crystallization and preliminary X-ray crystallographic studies of dehydroascorbate reductase (DHAR) from Oryza sativa L. japonica.


ABSTRACT: Dehydroascorbate reductase from Oryza sativa L. japonica (OsDHAR), a key enzyme in the regeneration of vitamin C, maintains reduced pools of ascorbic acid to detoxify reactive oxygen species. In previous studies, the overexpression of OsDHAR in transgenic rice increased grain yield and biomass as well as the amount of ascorbate, suggesting that ascorbate levels are directly associated with crop production in rice. Hence, it has been speculated that the increased level of antioxidants generated by OsDHAR protects rice from oxidative damage and increases the yield of rice grains. However, the crystal structure and detailed mechanisms of this important enzyme need to be further elucidated. In this study, recombinant OsDHAR protein was purified and crystallized using the sitting-drop vapour-diffusion method at pH 8.0 and 298?K. Plate-shaped crystals were obtained using 0.15?M potassium bromide, 30%(w/v) PEG MME 2000 as a precipitant, and the crystals diffracted to a resolution of 1.9?Å on beamline 5C at the Pohang Accelerator Laboratory. The X-ray diffraction data indicated that the crystal contained one OsDHAR molecule in the asymmetric unit and belonged to space group P2? with unit-cell parameters a=47.03, b=48.38, c=51.83?Å, ?=107.41°.

SUBMITTER: Do H 

PROVIDER: S-EPMC4051537 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature

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Crystallization and preliminary X-ray crystallographic studies of dehydroascorbate reductase (DHAR) from Oryza sativa L. japonica.

Do Hackwon H   Kim Il-Sup IS   Kim Young-Saeng YS   Shin Sun-Young SY   Kim Jin-Ju JJ   Mok Ji-Eun JE   Park Seong-Im SI   Wi Ah Ram AR   Park Hyun H   Kim Han-Woo HW   Yoon Ho-Sung HS   Lee Jun Hyuck JH  

Acta crystallographica. Section F, Structural biology communications 20140524 Pt 6


Dehydroascorbate reductase from Oryza sativa L. japonica (OsDHAR), a key enzyme in the regeneration of vitamin C, maintains reduced pools of ascorbic acid to detoxify reactive oxygen species. In previous studies, the overexpression of OsDHAR in transgenic rice increased grain yield and biomass as well as the amount of ascorbate, suggesting that ascorbate levels are directly associated with crop production in rice. Hence, it has been speculated that the increased level of antioxidants generated b  ...[more]

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