Crystallization and preliminary X-ray crystallographic study of the editing domain of Thermus thermophilus isoleucyl-tRNA synthetase complexed with pre- and post-transfer editing-substrate analogues.
The CP1 domain (the editing domain) of isoleucyl-tRNA synthetase (IleRS) hydrolyzes misactivated Val-AMP in pre-transfer editing and mischarged Val-tRNA(Ile) in post-transfer editing. The CP1 domain of Thermus thermophilus IleRS was expressed in isolation, purified and cocrystallized with Val-AMS (a Val-AMP analogue) and with Val-2AA (a Val-tRNA(Ile) analogue). Two different expression constructs were used for each cocrystallization. The complex crystals with Val-AMS belong to the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = b = 102.00, c = 84.88 A. The asymmetric unit contains two molecules of the CP1 domain, with a corresponding crystal volume per protein weight of 2.7 A3 Da(-1) and a solvent content of 53.5%. The complex crystals with Val-2AA belong to the tetragonal space group P4(1)22, with unit-cell parameters a = b = 72.59, c = 83.68 A. The asymmetric unit contains one molecule of the CP1 domain, with a corresponding crystal volume per protein weight of 2.8 A3 Da(-1) and a solvent content of 55.8%. Data sets diffracting to 1.7 A resolution were collected from each single crystal at 100 K.
Fukunaga
Ryuya
R
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Japan.
Yokoyama
Shigeyuki
S
D003460:Crystallization; D018360:Crystallography, X-Ray; D007533:Isoleucine-tRNA Ligase / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D017393:RNA Editing; D013696:Temperature; D016963:Thermus thermophilus / Q000201:enzymology*
D007533:Isoleucine-tRNA Ligase