Cloning, expression, crystallization and preliminary X-ray crystallographic analysis of glutamyl-tRNA synthetase (Xoo1504) from Xanthomonas oryzae pv. oryzae.
The gltX gene from Xanthomonas oryzae pv. oryzae (Xoo1504) encodes glutamyl-tRNA synthetase (GluRS), one of the most important enzymes involved in bacterial blight (BB), which causes huge production losses of rice worldwide. GluRS is a class I-type aminoacyl-tRNA synthetase (aaRS) that is primarily responsible for the glutamylation of tRNA(Glu). It plays an essential role in protein synthesis, as well as the regulation of cells, in all organisms. As it represents an important target for the development of new antibacterial drugs against BB, determination of the three-dimensional structure of GluRS is essential in order to understand its catalytic mechanism. In order to analyze its structure and function, the gltX gene was cloned and the GluRS enzyme was expressed, purified and then crystallized. A GluRS crystal belonging to the monoclinic space group C2 diffracted to 2.8 A resolution and had unit-cell parameters a = 186.8, b = 108.4, c = 166.1 A, beta = 96.3 degrees . The unit-cell volume of the crystal allowed the presence of six to eight monomers in the asymmetric unit, with a corresponding Matthews coefficient (V(M)) range of 2.70-2.02 A(3) Da(-1) and a solvent-content range of 54.5-39.3%.
Doan
Thanh Thi Ngoc
TT
Department of Advanced Technology Fusion, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul, Republic of Korea.
Natarajan
Sampath
S
Kim
Hyesoon
H
Ahn
Yeh-Jin
YJ
Kim
Jeong-Gu
JG
Lee
Byoung-Moo
BM
Kang
Lin-Woo
LW
D000890:Anti-Infective Agents / Q000737:chemistry; D002384:Catalysis; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D015195:Drug Design; D004591:Electrophoresis, Polyacrylamide Gel; D005975:Glutamate-tRNA Ligase / Q000737:chemistry*; D015233:Models, Statistical; D009584:Nitrogen / Q000737:chemistry; D012275:Oryza / Q000382:microbiology*; D010957:Plasmids / Q000378:metabolism; D046940:Synechococcus / Q000378:metabolism; D014961:X-Ray Diffraction; D014974:Xanthomonas / Q000378:metabolism*
D000890:Anti-Infective Agents; D005975:Glutamate-tRNA Ligase; D009584:Nitrogen