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 th...

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Published in:Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2005-04, Vol.61 (4), p.432-434
Main Authors: Higashi, Noriko, Matsuura, Takanori, Nakagawa, Atsushi, Ishikawa, Kazuhiko
Format: Article
Language:English
Subjects:
Alcohol Oxidoreductases - chemistry
archaea
Archaeal Proteins - chemistry
Cloning, Molecular - methods
CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY
CRYSTALLIZATION
Crystallization - methods
Crystallization Communications
CRYSTALS
DIFFRACTION
DIFFUSION
ESCHERICHIA COLI
Escherichia coli - genetics
hyperthermostability
MOLECULAR WEIGHT
Pyrococcus horikoshii
Pyrococcus horikoshii - chemistry
RESOLUTION
SOLVENTS
SPACE GROUPS
threonine dehydrogenase
Volatilization
X-Ray Diffraction
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Summary: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 Å resolution under cryogenic conditions. P. horikoshiil‐threonine dehydrogenase crystals belong to space group I4122, with unit‐cell parameters a = b = 143.84, c = 304.13 Å. The presence of three subunits of the enzyme per asymmetric unit was estimsted to give a Matthews coefficient (VM) of 3.5 Å3 Da−1 and a solvent content of 64.7%(v/v).
ISSN:1744-3091
1744-3091
DOI:10.1107/S174430910500881X