Open AccessCrystallization and preliminary X‐ray analysis of the yeast tRNA‐thiouridine modification protein 1 (Tum1p)
Author(s) -
Qiu Rui,
Wang Fengbin,
Liu Meiruo,
Yang Zhenxing,
Wu Tong,
Ji Chaoneng
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111024900
Subject(s) - crystallization , transfer rna , yeast , chemistry , x ray , crystallography , biochemistry , rna , physics , optics , organic chemistry , gene
Yeast tRNA‐thiouridine modification protein 1 (Tum1p), a crucial component of the Urm1 system, is believed to play important roles in protein urmylation and tRNA‐thiouridine modification. Previous studies have demonstrated that the conserved residue Cys259 in the C‐terminal rhodanese‐like domain of Tum1p is essential for these sulfur‐transfer activities. Here, recombinant Tum1p protein has been cloned and overexpressed in Escherichia coli strain BL21 (DE3). After purification, crystals of Tum1p were obtained by the hanging‐drop vapour‐diffusion method and diffracted to 1.9 Å resolution. The preliminary X‐ray data showed that the tetragonal Tum1p crystal belonged to space group I 4 1 , with unit‐cell parameters a = b = 120.94, c = 48.35 Å. The asymmetric unit of the crystal was assumed to contain one protein molecule, giving a Matthews coefficient of 2.41 Å 3 Da −1 and a solvent content of 49.0%.