Qiu Rui, Wang Fengbin, Liu Meiruo, Yang Zhenxing, Wu Tong, Ji Chaoneng
State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai, People's Republic of China.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Aug 1;67(Pt 8):953-5. doi: 10.1107/S1744309111024900. Epub 2011 Jul 27.
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 I4(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%.
酵母tRNA-硫代尿苷修饰蛋白1（Tum1p）是Urm1系统的关键组成部分，被认为在蛋白质 urmylation 和 tRNA-硫代尿苷修饰中发挥重要作用。先前的研究表明，Tum1p C 末端类硫氧还蛋白结构域中的保守残基 Cys259 对于这些硫转移活性至关重要。在此，重组 Tum1p 蛋白已在大肠杆菌 BL21（DE3）菌株中克隆并过量表达。纯化后，通过悬滴气相扩散法获得了 Tum1p 的晶体，并衍射至 1.9 Å 分辨率。初步的 X 射线数据表明，四方晶系的 Tum1p 晶体属于空间群 I4(1)，晶胞参数 a = b = 120.94，c = 48.35 Å。假定晶体的不对称单元包含一个蛋白质分子，马修斯系数为 2.41 Å(3) Da(-1)，溶剂含量为 49.0%。
