枯草芽孢杆菌色氨酰 - tRNA合成酶的结晶

Crystallization of Bacillus subtilis tryptophanyl-tRNA synthetase.

作者信息

Ala P, Xue H, Leung L, Xue Y Q, Wong J T, Yang D S

机构信息

Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.

出版信息

J Mol Biol. 1993 Apr 5;230(3):1089-90. doi: 10.1006/jmbi.1993.1223.

DOI:10.1006/jmbi.1993.1223

PMID:8478922

Abstract

Tryptophanyl-tRNA synthetase from Bacillus subtilis was overexpressed in Escherichia coli and was purified using column chromatography on DEAE-Sephacel and hydroxyapatite columns. Single crystals of the synthetase were grown by vapor diffusion at 4 degrees C from pH 5.5 solutions of polyethylene glycol 8000 containing magnesium ATP and L-tryptophan. The crystals diffracted to about 4.0 A resolution at -150 degrees C and appeared to belong to the orthorhombic space group P2(1)2(1)2 with unit cell dimensions: a = 143.6 A, b = 111.6 A, c = 50.6 A with one dimer in the asymmetric unit.

摘要

来自枯草芽孢杆菌的色氨酰 - tRNA合成酶在大肠杆菌中过表达，并通过在DEAE - Sephacel柱和羟基磷灰石柱上进行柱色谱法进行纯化。该合成酶的单晶在4℃下通过气相扩散法从含有镁ATP和L - 色氨酸的聚乙二醇8000的pH 5.5溶液中生长。这些晶体在-150℃下衍射至约4.0 Å分辨率，似乎属于正交晶系空间群P2(1)2(1)2，晶胞参数为：a = 143.6 Å，b = 111.6 Å，c = 50.6 Å，不对称单位中有一个二聚体。