Crystal structure analysis of a recombinant predicted acetamidase/formamidase from the thermophile Thermoanaerobacter tengcongensis.
The structure of acetamidase/formamidase (Amds/Fmds) from the archaeon Thermoanaerobacter tengcongensis has been determined by X-ray diffraction analysis using MAD data in a crystal of space group P2₁, with unit-cell parameters a = 41.23 (3), b = 152.88 (6), c = 100.26 (7) Å, β = 99.49 (3) ° and been refined to a crystallographic R-factor of 17.4% and R-free of 23.7%. It contains two dimers in one asymmetric unit, in which native Amds/Fmds (TE19) contains of the 32 kDa native protein. The final model consists of 4 monomer (299 amino acids residues with additional 2 expression tag amino acids residues), 5 Ca²⁺, 4 Zn²⁺ and 853 water molecules. The monomer is composed by the following: an N-domain which is featuring by three-layers β/β/β; a prominent excursion between N-terminal end of strand β₇ and β₁₁, which contains four-stranded antiparallel β sheet; an C-domain which is formed by the last 82 amino acid residues with the feature of mixed α/β structure. The protein contains ion-pair Ca²⁺-Zn²⁺. The portion of three-layer β/β/β along with the loops provides four protein ligands to the tightly bound Ca²⁺, three water molecules complete the coordination; and provides five protein ligands to the tightly bound Zn²⁺, one water molecule complete the coordination.
State Key Laboratory for Structural Chemistry of Unstable and Stable Species, College of Chemistry, Peking University, Beijing, People's Republic of China. qianmx@pku.edu.cn
Minxie
M
Qian
Qichen
Q
Huang
Guangteng
G
Wu
Luhua
L
Lai
Youqi
Y
Tang
Jianfeng
J
Pei
Masami
M
Kusunoki
D000581:Amidohydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D019281:Dimerization; D008024:Ligands; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012645:Selenomethionine / Q000737:chemistry / Q000378:metabolism; D046968:Thermoanaerobacter / Q000201:enzymology*; D014867:Water / Q000737:chemistry / Q000378:metabolism; D015032:Zinc / Q000737:chemistry / Q000378:metabolism
D001426:Bacterial Proteins; D008024:Ligands; D011994:Recombinant Proteins; D014867:Water; D012645:Selenomethionine; D000581:Amidohydrolases; C021445:acetamidase; C101846:formamidase; D015032:Zinc; D002118:Calcium