Structural understanding of the recycling of oxidized ascorbate by dehydroascorbate reductase (OsDHAR) from Oryza sativa L. japonica

Scientific Reports
Hackwon DoJun Hyuck Lee

Abstract

Dehydroascorbate reductase (DHAR) is a key enzyme involved in the recycling of ascorbate, which catalyses the glutathione (GSH)-dependent reduction of oxidized ascorbate (dehydroascorbate, DHA). As a result, DHAR regenerates a pool of reduced ascorbate and detoxifies reactive oxygen species (ROS). In previous experiments involving transgenic rice, we observed that overexpression of DHAR enhanced grain yield and biomass. Since the structure of DHAR is not available, the enzymatic mechanism is not well-understood and remains poorly characterized. To elucidate the molecular basis of DHAR catalysis, we determined the crystal structures of DHAR from Oryza sativa L. japonica (OsDHAR) in the native, ascorbate-bound, and GSH-bound forms and refined their resolutions to 1.9, 1.7, and 1.7 Å, respectively. These complex structures provide the first information regarding the location of the ascorbate and GSH binding sites and their interacting residues. The location of the ascorbate-binding site overlaps with the GSH-binding site, suggesting a ping-pong kinetic mechanism for electron transfer at the common Cys20 active site. Our structural information and mutagenesis data provide useful insights into the reaction mechanism of OsDHAR agains...Continue Reading

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Citations

Jan 18, 2017·Chemical Communications : Chem Comm·Jun Ohata, Zachary T Ball
Feb 29, 2020·Journal of Experimental Botany·Haiyan DingShengchun Li
Nov 20, 2019·Toxins·Lakshmipriya PerincherryŁukasz Stępień
Jan 16, 2020·International Journal of Molecular Sciences·Sara HamzelouPaul A Haynes
Aug 2, 2020·Free Radical Biology & Medicine·David NjusH Bernhard Schlegel
Apr 14, 2016·Biochemical and Biophysical Research Communications·Bhaba Krishna DasArulandu Arockiasamy
May 20, 2021·Plant Physiology and Biochemistry : PPB·Walid DerbaliHans-Werner Koyro
Jul 3, 2021·Plants·Inês LeitãoMiguel P Mourato
Apr 14, 2018·Journal of Medicinal Chemistry·Yiyue XieJonathan B Baell

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Methods Mentioned

BETA
transgenic
AsA
protein folding
PCR
gel filtration
X-ray
circular dichroism

Software Mentioned

MODEL
REFMAC5
HKL
COOT
MOLREP
MolProbity
Phenix
SWISS
2000

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