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嗜热栖热菌甘氨酰-tRNA合成酶的结晶及初步晶体学数据。

Crystallisation of the glycyl-tRNA synthetase from Thermus thermophilus and initial crystallographic data.

作者信息

Logan D T, Cura V, Touzel J P, Kern D, Moras D

机构信息

UPR de Biologie Structurale, Strasbourg, France.

出版信息

J Mol Biol. 1994 Sep 2;241(5):732-5. doi: 10.1006/jmbi.1994.1547.

Abstract

The glycyl-tRNA synthetase from Thermus thermophilus is a dimer of molecular mass 115 kDa, which has been crystallised using the vapour diffusion method from 5 to 7% polyethylene glycol 6000, 0.8 to 1.4 M NaCl at protein concentrations of 2 to 8 mg/ml. Nucleation is carried out at 4 degrees C and crystals are subsequently transferred to 15 degrees C to maximise growth. Crystals are truncated rhombohedra measuring on average 0.4 mm x 0.4 mm x 0.2 mm, which appear within a few days and reach full size in one to two months. GlyRS crystallises in two closely related space groups, P2(1)2(1)2(1) and C2,2,2(1), both with the same cell a = 125 A, b = 254 A, c = 104 A. Crystal packing in P2(1)2(1)2(1) is strongly C-centred. The crystals have VM = 3.6 A3/Da and a solvent content of 61%, with one dimer in the asymmetric unit in C2,2,2(1) and two dimers in P2(1)2(1)2(1). The best native data extend to 2.9 A in C2,2,2(1) and are 90.6% complete with an R-factor between symmetry-related reflections of 10.0%. The structure has been solved by multiple isomorphous replacement and model building is in progress.

摘要

嗜热栖热菌的甘氨酰 - tRNA合成酶是一种分子量为115 kDa的二聚体,已使用气相扩散法在5%至7%的聚乙二醇6000、0.8至1.4 M氯化钠中,蛋白质浓度为2至8 mg/ml的条件下进行结晶。成核在4℃下进行,随后将晶体转移至15℃以促进生长。晶体为截顶菱面体,平均尺寸为0.4 mm×0.4 mm×0.2 mm,在几天内出现,并在一到两个月内长到完整尺寸。甘氨酰 - tRNA合成酶(GlyRS)在两个紧密相关的空间群P2(1)2(1)2(1)和C2,2,2(1)中结晶,两者的晶胞参数相同,a = 125 Å,b = 254 Å,c = 104 Å。P2(1)2(1)2(1)中的晶体堆积以C心为主。晶体的VM = 3.6 Å3/Da,溶剂含量为61%,在C2,2,2(1)的不对称单元中有一个二聚体,在P2(1)2(1)2(1)中有两个二聚体。在C2,2,2(1)中,最佳的天然数据延伸至2.9 Å,完整性为90.6%,对称相关反射之间的R因子为10.0%。该结构已通过多对同晶置换法解析,模型构建正在进行中。

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