哺乳动物线粒体丝氨酰-tRNA合成酶的结晶及初步X射线衍射研究
Crystallization and preliminary X-ray diffraction study of mammalian mitochondrial seryl-tRNA synthetase.
作者信息
Chimnaronk Sarin, Jeppesen Mads Gravers, Shimada Nobukazu, Suzuki Tsutomu, Nyborg Jens, Watanabe Kimitsuna
机构信息
Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan.
出版信息
Acta Crystallogr D Biol Crystallogr. 2004 Jul;60(Pt 7):1319-22. doi: 10.1107/S0907444904011217. Epub 2004 Jun 22.
The mitochondrial seryl-tRNA synthetase (mt SerRS) from Bos taurus was overexpressed in Escherichia coli and crystallized using the sitting-drop vapour-diffusion method. Crystals grew in a very narrow range of conditions using PEG 8000 as precipitant at room temperature. An appropriate concentration of lithium sulfate was critical for crystal nucleation. Crystals diffracted well beyond a resolution of 1.6 A and were found to belong to the orthorhombic space group C222(1), with unit-cell parameters a = 79.89, b = 230.42, c = 135.60 A. There is one dimer (M(r) approximately 113 kDa) in the asymmetric unit, with a solvent content of 55%. Efforts to solve the phase problem by molecular replacement are under way.
来自牛的线粒体丝氨酰 - tRNA合成酶（mt SerRS）在大肠杆菌中过量表达，并采用坐滴气相扩散法进行结晶。在室温下使用聚乙二醇8000作为沉淀剂，晶体在非常狭窄的条件范围内生长。适当浓度的硫酸锂对晶体成核至关重要。晶体衍射分辨率超过1.6 Å，属于正交晶系空间群C222(1)，晶胞参数a = 79.89，b = 230.42，c = 135.60 Å。不对称单元中有一个二聚体（相对分子质量约为113 kDa），溶剂含量为55%。目前正在通过分子置换法解决相位问题。
Zotero 插件