Crystallization and preliminary x-ray diffraction analysis of L-threonine dehydrogenase (TDH) from the hyperthermophilic archaeon Thermococcus kodakaraensis

• Main Authors: Bowyer, A. (Author), Mikolajek, H. (Author), Wright, J.N (Author), Coker, A. (Author), Erskine, P.T (Author), Cooper, J.B (Author), Bashir, Q. (Author), Rashid, N. (Author), Jamil, F. (Author), Akhtar, M. (Author)
• Format: Article
• Language: English
• Published: 2008-08-20.
• Subjects: Article
• Online Access: Get fulltext

The enzyme L-threonine dehydrogenase catalyses the NAD(+)-dependent conversion of L-threonine to 2-amino-3-ketobutyrate, which is the first reaction of a two-step biochemical pathway involved in the metabolism of threonine to glycine. Here, the crystallization and preliminary crystallographic analysis of L-threonine dehydrogenase (Tk-TDH) from the hyperthermophilic organism Thermococcus kodakaraensis KOD1 is reported. This threonine dehydrogenase consists of 350 amino acids, with a molecular weight of 38 kDa, and was prepared using an Escherichia coli expression system. The purified native protein was crystallized using the hanging-drop vapour-diffusion method and crystals grew in the tetragonal space group P4(3)2(1)2, with unit-cell parameters a = b = 124.5, c = 271.1 A. Diffraction data were collected to 2.6 A resolution and preliminary analysis indicates that there are four molecules in the asymmetric unit of the crystal.