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.
KeywordsAcetamidase/formamidase Structural genomics Crystal structure
- T. tengcongensis
Multi-wavelength anomalous dispersion
Our appreciation to Professor Ruseng Chen (Institute of Biophysics, Academia Sinica) for providing the genomic DNA of T. tengcongensis also to Drs Soichi Wakatsuki and Noriyuki Igarashi for their kind helps with data collection at the Photon Factory. This work has been supported by grants from “Structural Genomics” of the High Technology Development Program of “863 Project of China” to QH and partially by Grants-in-Aid (Nos. 10558109 and 12480181) from the Japan Society for the Promotion of Science to MK.
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