Purification, characterization and preliminary X-ray crystallographic studies of monodehydroascorbate reductase from Oryza sativa L. japonica.
Acta Crystallogr F Struct Biol Commun
; 70(Pt 9): 1244-8, 2014 Sep.
Article
em En
| MEDLINE
| ID: mdl-25195901
ABSTRACT
Monodehydroascorbate reductase (MDHAR; EC 1.6.5.4) is a key enzyme in the reactive oxygen species (ROS) detoxification system of plants. The participation of MDHAR in ascorbate (AsA) recycling in the ascorbate-glutathione cycle is important in the acquired tolerance of crop plants to abiotic environmental stresses. Thus, MDHAR represents a strategic target protein for the improvement of crop yields. Although physiological studies have intensively characterized MDHAR, a structure-based functional analysis is not available. Here, a cytosolic MDHAR (OsMDHAR) derived from Oryza sativa L. japonica was expressed using Escherichia coli strain NiCo21 (DE3) and purified. The purified OsMDHAR showed specific enzyme activity (approximately 380â
U per milligram of protein) and was crystallized using the hanging-drop vapour-diffusion method at pH 8.0 and 298â
K. The crystal diffracted to 1.9â
Å resolution and contained one molecule in the asymmetric unit (the Matthews coefficient VM is 1.98â
Å(3)â
Da(-1), corresponding to a solvent content of 38.06%) in space group P41212 with unit-cell parameters a = b = 81.89, c = 120.4â
Å. The phase of the OsMDHAR structure was resolved by the molecular-replacement method using a ferredoxin reductase from Acidovorax sp. strain KKS102 (PDB entry 4h4q) as a model.
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MEDLINE
Assunto principal:
Oryza
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Cristalografia por Raios X
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NADH NADPH Oxirredutases
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article