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

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Title
Crystallization and preliminary X-ray crystallographic studies of dehydroascorbate reductase (DHAR) from Oryza sativa L. japonica
Other Titles
벼유래 dehydroascorbate reductase (DHAR) 효소의 결정화 및 삼차구조해석을 위한 X-선 회절데이타 분석
Authors
Il-Sup Kim
Lee, Jun Hyuck
Yoon, Ho-Sung
Kim, Han-Woo
Park, Hyun
Wi, Ah Ram
Seong-Im Park
Ji-Eun Mok
Jin-Ju Kim
Sun-Young Shin
Do, Hackwon
Young-Saeng Kim
Keywords
Oryza sativa L. japonica; X-ray crystallography; ascorbate; dehydroascorbate reductase
Issue Date
2014
Citation
Il-Sup Kim, et al. 2014. "Crystallization and preliminary X-ray crystallographic studies of dehydroascorbate reductase (DHAR) from Oryza sativa L. japonica". ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS, 70(6): 781-785.
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 (ROS). 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, the increased level of antioxidants generated by OsDHAR has been speculated to protect rice from oxidative damage and increase the yield of germination. 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 vapor diffusion method at pH 8.0 and 298 K. Plate-shape crystals were obtained using 0.15 M potassium bromide and 30% (w/v) PEG MME 2000 as a precipitant, and the crystals diffracted to a resolution of 1.9 A at beamline 5C of the Pohang Accelerator Laboratory. The X-ray diffraction data indicated that the crystal contained one OsDHAR molecule in the asymmetric unit with the space group P21 (unit-cell parameters, a = 47.03, b = 48.38, c = 51.83 A, and β = 107.41°).
URI
http://repository.kopri.re.kr/handle/201206/7268
DOI
http://dx.doi.org/10.1107/S2053230X14009133
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