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酵母精氨酰 - tRNA合成酶 - 酵母tRNAArg复合物的结晶及初步X射线晶体学分析

Crystallization and preliminary X-ray crystallographic analysis of yeast arginyl-tRNA synthetase-yeast tRNAArg complexes.

作者信息

Delagoutte B, Keith G, Moras D, Cavarelli J

机构信息

UPR 9004 Biologie Structurale, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, BP 163, 67404 Illkirch CEDEX, France.

出版信息

Acta Crystallogr D Biol Crystallogr. 2000 Apr;56(Pt 4):492-4. doi: 10.1107/s0907444900001700.

Abstract

Three different crystal forms of complexes between arginyl-tRNA synthetase from the yeast Saccharomyces cerevisae (yArgRS) and the yeast second major tRNA(Arg) (tRNA(Arg)(ICG)) isoacceptor have been crystallized by the hanging-drop vapour-diffusion method in the presence of ammonium sulfate. Crystal form II, which diffracts beyond 2.2 A resolution at the European Synchrotron Radiation Facility ID14-4 beamline, belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 129.64, b = 107.47, c = 71. 38 A. This crystal form presents the highest resolution obtained for an active form of an aminoacyl-tRNA synthetase-tRNA complex. The estimated V(m) of 2.6 A(3) Da(-1) indicates one molecule of complex in the asymmetric unit. The three crystal forms were solved by the molecular-replacement method using the coordinates of the free yArgRS.

摘要

利用悬滴气相扩散法,在硫酸铵存在的情况下,已使来自酿酒酵母的精氨酰 - tRNA合成酶(yArgRS)与酵母第二主要tRNA(Arg)(tRNA(Arg)(ICG))同功受体之间的复合物形成了三种不同的晶体形式。晶体形式II在欧洲同步辐射装置ID14 - 4光束线处衍射分辨率超过2.2 Å,属于正交空间群P2(1)2(1)2,晶胞参数a = 129.64,b = 107.47,c = 71.38 Å。这种晶体形式是氨基酰 - tRNA合成酶 - tRNA复合物活性形式所获得的最高分辨率。估计的V(m)为2.6 ų Da⁻¹,表明不对称单位中有一个复合物分子。通过使用游离yArgRS的坐标,采用分子置换法解析了这三种晶体形式。

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