Purification, crystallization and preliminary crystallographic data for rat cytosolic selenocysteine 498 to cysteine mutant thioredoxin reductase

Citation
Lw. Zhong et al., Purification, crystallization and preliminary crystallographic data for rat cytosolic selenocysteine 498 to cysteine mutant thioredoxin reductase, ACT CRYST D, 56, 2000, pp. 1191-1193
Citations number
19
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Chemistry & Analysis
Journal title
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
ISSN journal
0907-4449 → ACNP
Volume
56
Year of publication
2000
Part
9
Pages
1191 - 1193
Database
ISI
SICI code
0907-4449(200009)56:<1191:PCAPCD>2.0.ZU;2-W
Abstract
Mammalian cytosolic thioredoxin reductase is a homodimer of 55 kDa subunit containing an essential penultimate selenocysteine residue. An active analo gue of the rat enzyme in which cysteine replaces selenocysteine has been ex pressed in Escherichia coli cells at high levels and purified to homogeneit y. The pure enzyme contains one FAD per subunit and shows spectral properti es identical to that of the wild-type thioredoxin reductase. The isolated e nzyme in its oxidized and reduced forms or the enzyme complexed with NADP() was crystallized by the hanging-drop vapour-diffusion method. The diffrac tion pattern extends to 3 Angstrom resolution. The crystals are monoclinic, space group P2(1), with unit-cell parameters a = 78.9, b = 140.5, c = 170. 8 Angstrom, alpha = 94.6 degrees. There are three dimeric molecules in the asymmetric unit.