Open AccessCrystallization and preliminary X‐ray crystallographic study of the wild type and two mutants of the CP1 hydrolytic domain from Aquifex aeolicus leucyl‐tRNA synthetase
Author(s) -
Cura Vincent,
Olieric Natacha,
Guichard Alexandre,
Wang EnDuo,
Moras Dino,
Eriani Gilbert,
Cavarelli Jean
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105028460
Subject(s) - aquifex aeolicus , alanine , crystallography , crystallization , chemistry , threonine , serine , mutant , stereochemistry , hydrolysis , amino acid , biochemistry , enzyme , organic chemistry , escherichia coli , gene
The editing or hydrolytic CP1 domain of leucyl‐tRNA synthetase (LeuRS) hydrolyses several misactivated amino acids. The CP1 domain of Aquifex aeolicus LeuRS was expressed, purified and crystallized by the hanging‐drop vapour‐diffusion method using ammonium sulfate as precipitant. Crystals belong to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 38.8, b = 98.4, c = 116.7 Å. Crystals diffract to beyond 1.8 Å resolution and contain two monomers in the asymmetric unit. Two CP1 mutants in which a conserved threonine residue essential for the fidelity of the hydrolytic pathway is mutated to alanine or glutamic acid have also been expressed and crystallized. Crystals of the two CP1 mutants are isomorphs of the wild type and diffract to beyond 1.9 Å resolution. All structures were solved by molecular‐replacement techniques.