Protein preparation, crystallization and preliminary X-ray analysis of imidazolonepropionase from Bacillus subtilis.
Imidazolonepropionase (EC 3.5.2.7) is the third enzyme of the histidine degradation pathway that has been conserved from bacteria to eukaryotes. The enzyme is the only one with unknown three-dimensional structure in this pathway. In this work, Bacillus subtilis imidazolonepropionase (HutI) was expressed in E. coli and purified to homogeneity. After thrombin digestion, high quality crystals were obtained by hanging-drop vapor diffusion method. The best crystal diffracted to 2.0 A and belonged to the space group P2(1) with unit-cell parameters a = 57.73 A, b = 106.34 A, c = 66.47 A, beta = 89.93 degrees .
Yu
Yamei
Y
The National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, PR China.
Li
Lanfen
L
Zheng
Xiaofeng
X
Liang
Yu-He
YH
Su
Xiao-Dong
XD
D000581:Amidohydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray; D005798:Genes, Bacterial; D006639:Histidine / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006639:Histidine; D000581:Amidohydrolases; C092896:imidazolonepropionase, Bacillus subtilis