Crystallization and preliminary crystallographic study of a recombinant predicted acetamidase/formamidase from the thermophile Thermoanaerobacter tengcongensis.
No crystal structures are yet available for homologues of a predicted acetamidase/formamidase (Amds/Fmds) from the archaeon Thermoanaerobacter tengcongensis. The Amds/Fmds gene was cloned and expressed as a soluble protein in Escherichia coli. Native Amds/Fmds and its SeMet-substituted form were purified and crystallized by vapour diffusion in hanging drops at 296 K. The native crystals, which were grown in PEG 8000, belong to the monoclinic space group P2(1), with unit-cell parameters a = 41.23 (3), b = 152.88 (6), c = 100.26 (7) A, beta = 99.49 (3) degrees. The diffraction data were collected to 2.00 A resolution using synchrotron radiation. Based on a predicted solvent content of 50%, a Matthews coefficient of 2.44 A3 Da(-1) and two main peaks in the self-rotation function, the asymmetric unit is predicted to contain two dimers of the 32 kDa native protein. MAD data were collected for the SeMet protein, but the corresponding crystals display different unit-cell parameters and appear to contain four dimers in the asymmetric unit.
Wu
Guangteng
G
State Key Laboratory for Structural Chemistry of Unstable and Stable Species, College of Chemistry, Peking University, Beijing 100871, People's Republic of China.
Huang
Qichen
Q
Tang
Youqi
Y
Unno
Hideaki
H
Kusunoki
Masami
M
D000581:Amidohydrolases / Q000737:chemistry* / Q000302:isolation & purification; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology; D009097:Multienzyme Complexes / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012645:Selenomethionine; D017356:Synchrotrons; D046968:Thermoanaerobacter / Q000201:enzymology*
D001426:Bacterial Proteins; D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D012645:Selenomethionine; D000581:Amidohydrolases; C021445:acetamidase; C101846:formamidase