Preparation, crystallization and preliminary crystallographic analysis of old yellow enzyme from Trypanosoma cruzi.
Old yellow enzyme (OYE) is an NADPH oxidoreductase that contains a flavin mononucleotide as a prosthetic group. The OYE from Trypanosoma cruzi, which produces prostaglandin F(2alpha), a potent mediator of various physiological and pathological processes, from prostaglandin H2. The protein was recombinantly expressed and purified from Escherichia coli and was crystallized using the hanging-drop vapour-diffusion method. The crystal belongs to the monoclinic space group P2(1), with unit-cell parameters a = 56.3, b = 78.8, c = 78.8 A, beta = 93.4 degrees and two molecules per asymmetric unit. The crystals were suitable for X-ray crystallographic studies and diffracted to 1.70 A resolution. A Patterson search method is in progress using the structure of OYE from Pseudomonas putida as a starting model.
Sugiyama
Shigeru
S
Maruwa Foods Co. Ltd, Tsutsui-cho 170-1, Yamatokoriyama, Nara 639-1123, Japan.
Tokuoka
Keiji
K
Uchiyama
Nahoko
N
Okamoto
Naoki
N
Okano
Yousuke
Y
Matsumura
Hiroyoshi
H
Inaka
Koji
K
Urade
Yoshihiro
Y
Inoue
Tsuyoshi
T
D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D009252:NADPH Dehydrogenase / Q000032:analysis* / Q000737:chemistry*; D014349:Trypanosoma cruzi / Q000201:enzymology*
D009252:NADPH Dehydrogenase