Expression, purification, crystallization and preliminary crystallographic study of a potential metal-dependent hydrolase with cyclase activity from Thermoanaerobacter tengcongensis.
The putative metal-dependent hydrolase gene TTE1006 from Thermoanaerobacter tengcongensis strain MB4T (T = type strain; Genbank accession No. AE008691) was heterologously expressed in Escherichia coli. The 205-amino-acid gene product was purified and crystallized. The crystal used for data collection belongs to space group P2(1), with unit-cell parameters a = 85.2, b = 62.1, c = 172.4 A, beta = 104.2 degrees. Using a synchrotron-radiation source, the resolution limit of the data reached 1.87 A. Eight molecules were estimated to be present in the asymmetric unit, with a solvent content of 48%. Structure determination is ongoing using the multiple-wavelength anomalous diffraction (MAD) method and also the molecular-replacement (MR) method.
Liu
Sen
S
State Key Laboratory for Structural Chemistry of Unstable and Stable Species, College of Chemistry, Peking University, Beijing 100871, People's Republic of China.
Wu
Guangteng
G
Huang
Qichen
Q
Lai
Luhua
L
Tang
Youqi
Y
Unno
Hideaki
H
Kusunoki
Masami
M
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D003460:Crystallization; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D017356:Synchrotrons; D046968:Thermoanaerobacter / Q000201:enzymology*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006867:Hydrolases