Crystallization and preliminary crystallographic analysis of monodehydroascorbate radical reductase from cucumber.
Monodehydroascorbate (MDA) radical reductase (EC 1.6.5.4) is an FAD enzyme that catalyzes the univalent reduction of MDA radical to ascorbate using NAD(P)H as an electron donor. The recombinant MDA reductase from cucumber was crystallized using polyethylene glycol 6000 as a precipitant. The crystals belong to space group P2(1), with unit-cell parameters a = 60.8, b = 138.6, c = 61.7 A, beta = 114.5 degrees, and contained two molecules per asymmetric unit. The Matthews coefficient (VM) and the solvent content are 2.46 A3 Da(-1) and 50.0%, respectively. Diffraction data were collected to a resolution of 2.4 A at 100 K using Cu Kalpha radiation with a multi-wire area detector and gave a data set with an overall Rsym of 10.0% and a completeness of 92.5%.
Laboratory of Chemistry of Biological Function, Graduate School of Agriculture, Kyoto Prefectural University, Shimogamo, Sakyo, 606-8522, Japan.
Satoshi
S
Sano
You-Na
YN
Kang
Hiroko
H
Shigemizu
Nobuhiko
N
Morishita
Hye-Jin
HJ
Yoon
Kazumi
K
Saito
Kozi
K
Asada
Bunzo
B
Mikami
D003460:Crystallization; D018360:Crystallography, X-Ray; D018553:Cucumis sativus / Q000201:enzymology*; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry*
D009247:NADH, NADPH Oxidoreductases; C023927:monodehydroascorbate reductase (NADH)