Crystallization of leucyl-tRNA synthetase complexed with tRNALeu from the archaeon Pyrococcus horikoshii

被引：6

作者：

Fukunaga, R ^{[1
]}

Ishitani, R ^{[1
]}

Nureki, O ^{[1
]}

Yokoyama, S ^{[1
]}

机构：

[1] Univ Tokyo, Grad Sch Sci, Dept Biophys & Biochem, Tokyo, Japan

来源：

ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | 2005年 / 61卷

关键词：

D O I：

10.1107/S1744309104021827

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

All five tRNA (Leu) isoacceptors from the archaeon Pyrococcus horikoshii have been transcribed in vitro and purified. The leucyl-tRNA synthetase (LeuRS) from P. horikoshii was overexpressed in Escherichia coli and purified, and cocrystallizations with each of the tRNA(Leu) isoacceptors were attempted. Cocrystals were obtained by the hanging-drop vapour-diffusion method, but only when the tRNA(Leu) isoacceptor with the anticodon CAA was used. Electrophoretic analyses revealed that the crystals contain both LeuRS and tRNA(Leu), suggesting that they are LeuRS-tRNA(Leu) complex crystals. A data set diffracting to 3.3 A resolution was collected from a single crystal at 100 K. The crystal belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 118.18, b = 120.55, c = 231.13 angstrom. The asymmetric unit is expected to contain two complexes of LeuRS-tRNA(Leu), with a corresponding crystal volume per protein weight of 2.9 angstrom(3) Da(-1) and a solvent content of 57.3%.

引用

页码：30 / 32

页数：3

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