Crystallization and preliminary X-ray analysis of hyperthermophilic L-threonine dehydrogenase from the archaeon Pyrococcus horikoshii.
Recombinant L-threonine dehydrogenase from the hyperthermophilic archaeon Pyrococcus horikoshii was prepared using an Escherichia coli expression system. The hyperthermostable L-threonine dehydrogenase consists of 348 amino acids with a molecular weight of 37.7 kDa. The enzyme was crystallized by the hanging-drop vapour-diffusion method at 277 K and preliminary X-ray crystallographic analysis was carried out. Diffraction data were collected to 2.20 A resolution under cryogenic conditions. P. horikoshii L-threonine dehydrogenase crystals belong to space group I4(1)22, with unit-cell parameters a = b = 143.84, c = 304.13 A. The presence of three subunits of the enzyme per asymmetric unit was estimated to give a Matthews coefficient (VM) of 3.5 A3 Da(-1) and a solvent content of 64.7%(v/v).
Higashi
Noriko
N
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST Kansai), 1-8-31 Midorigaoka, Ikeda, Osaka 563-8577, Japan.
Matsuura
Takanori
T
Nakagawa
Atsushi
A
Ishikawa
Kazuhiko
K
D000429:Alcohol Oxidoreductases / Q000737:chemistry*; D019843:Archaeal Proteins / Q000737:chemistry; D003001:Cloning, Molecular / Q000379:methods; D003460:Crystallization / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D043902:Pyrococcus horikoshii / Q000737:chemistry*; D014835:Volatilization; D014961:X-Ray Diffraction
D019843:Archaeal Proteins; D000429:Alcohol Oxidoreductases; C023159:L-threonine 3-dehydrogenase