Purification, crystallization and preliminary X-ray crystallographic analysis of branched-chain aminotransferase from Deinococcus radiodurans.
The branched-chain amino-acid aminotransferase (BCAT), which requires pyridoxal 5'-phosphate (PLP) as a cofactor, is a key enzyme in the biosynthetic pathway of the hydrophobic amino acids leucine, isoleucine and valine. DrBCAT from Deinococcus radiodurans, which has a molecular weight of 40.9 kDa, was crystallized using the hanging-drop vapour-diffusion method. According to X-ray diffraction data to 2.50 A resolution from a DrBCAT crystal, the crystal belongs to space group P2(1)2(1)2(1), with unit-cell parameters a = 56.37, b = 90.70, c = 155.47 A. Preliminary analysis indicates the presence of two DrBCAT molecules in the asymmetric unit, with a solvent content of 47.52%.
Chen
Chung-Der
CD
Department of Physics, National Tsing-Hua University, Hsinchu 30013, Taiwan.
Huang
Tien-Feng
TF
Lin
Chih-Hao
CH
Guan
Hong-Hsiang
HH
Hsieh
Yin-Cheng
YC
Lin
Yi-Hung
YH
Huang
Yen-Chieh
YC
Liu
Ming-Yih
MY
Chang
Wen-Chang
WC
Chen
Chun-Jung
CJ
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D003460:Crystallization; D018360:Crystallography, X-Ray; D034301:Deinococcus / Q000201:enzymology*; D007532:Isoleucine / Q000737:chemistry; D007930:Leucine / Q000737:chemistry; D000637:Transaminases / Q000737:chemistry / Q000302:isolation & purification*; D014633:Valine / Q000737:chemistry
D001426:Bacterial Proteins; D007532:Isoleucine; D000637:Transaminases; C019798:branched-chain-amino-acid transaminase; D007930:Leucine; D014633:Valine