title: Crystallization and preliminary X-ray crystallographic study of the wild type and two mutants of the CP1 hydrolytic domain from Aquifex aeolicus leucyl-tRNA synthetase.
abstract: 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 P2(1)2(1)2(1), with unit-cell parameters a = 38.8, b = 98.4, c = 116.7 A. Crystals diffract to beyond 1.8 A 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 A resolution. All structures were solved by molecular-replacement techniques.
authors.affiliation: Département de Biologie et Génomique Structurales, UMR 7104, Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP Strasbourg, 1 Rue Laurent Fries, 67404 Illkirch, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Cura
authors.forename: Natacha
authors.initials: N
authors.lastname: Olieric
authors.forename: Alexandre
authors.initials: A
authors.lastname: Guichard
authors.forename: En-Duo
authors.initials: ED
authors.lastname: Wang
authors.forename: Dino
authors.initials: D
authors.lastname: Moras
authors.forename: Gilbert
authors.initials: G
authors.lastname: Eriani
authors.forename: Jean
authors.initials: J
authors.lastname: Cavarelli
mesh_terms: D000409:Alanine / Q000737:chemistry; D001419:Bacteria / Q000201:enzymology*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D018698:Glutamic Acid / Q000737:chemistry; D006868:Hydrolysis; D007935:Leucine-tRNA Ligase / Q000737:chemistry*; D009154:Mutation; D011487:Protein Conformation; D020033:Protein Isoforms; D017434:Protein Structure, Tertiary; D014961:X-Ray Diffraction
chemical_list: D020033:Protein Isoforms; D018698:Glutamic Acid; D007935:Leucine-tRNA Ligase; D000409:Alanine