Crystallization and preliminary crystallographic analysis of Thermus thermophilus leucyl-tRNA synthetase and its complexes with leucine and a non-hydrolysable leucyl-adenylate analogue.
Leucyl-tRNA synthetase from Thermus thermophilus (LeuRSTT) is the first LeuRS to be crystallized. Two crystal forms of the native enzyme have been obtained using the hanging-drop vapour-diffusion method with ammonium sulfate as a precipitant. Crystals of the first form belong to space group I422 and have unit-cell parameters a = b = 312.4, c = 100.4 A. They diffract anisotropically to 3.5 A resolution in the c-axis direction and to only 6 A resolution in the perpendicular direction. Crystals of the second form, which can be obtained native or with leucine or a leucyl-adenylate analogue bound, belong to space group C222(1) and have unit-cell parameters a = 102. 4, b = 154.1, c = 174.3 A. They diffract to 1.9 A resolution and contain one monomer in the asymmetric unit. Selenomethionated LeuRSTT has been produced and crystals of the second form suitable for MAD analysis have been grown.
Yaremchuk
A
A
EMBL, Grenoble Outstation, c/o ILL, 6 Rue Jules Horowitz, F-38042 Grenoble CEDEX 9, France.
Cusack
S
S
Gudzera
O
O
Grøtli
M
M
Tukalo
M
M
D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods; D007930:Leucine / Q000031:analogs & derivatives* / Q000378:metabolism*; D007935:Leucine-tRNA Ligase / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D012645:Selenomethionine / Q000378:metabolism; D016963:Thermus thermophilus / Q000201:enzymology*
D012645:Selenomethionine; D007935:Leucine-tRNA Ligase; D007930:Leucine