Open AccessCrystallization and preliminary X‐ray analysis of d ‐2‐hydroxyacid dehydrogenase from Haloferax mediterranei
Author(s) -
Domenech J.,
Baker P. J.,
Sedelnikova S. E.,
Rodgers H. F.,
Rice D. W.,
Ferrer J.
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910900863x
Subject(s) - triclinic crystal system , monoclinic crystal system , crystallography , crystallization , chemistry , ammonium sulfate , resolution (logic) , ternary operation , crystal (programming language) , crystal structure , chromatography , organic chemistry , computer science , programming language , artificial intelligence
d ‐2‐Hydroxyacid dehydrogenase (D2‐HDH) from Haloferax mediterranei has been overexpressed in Escherichia coli , solubilized in 8 M urea and refolded by rapid dilution. The protein was purified and crystallized by the hanging‐drop vapour‐diffusion method using ammonium sulfate or PEG 3350 as precipitant. Two crystal forms representing the free enzyme and the nonproductive ternary complex with α‐ketohexanoic acid and NAD + grew under these conditions. Crystals of form I diffracted to beyond 3.0 Å resolution and belonged to the monoclinic space group P 2 1 , with unit‐cell parameters a = 66.0, b = 119.6, c = 86.2 Å, β = 96.3°. Crystals of form II diffracted to beyond 2.0 Å resolution and belonged to the triclinic space group P 1, with unit‐cell parameters a = 66.5, b = 75.2, c = 77.6 Å, α = 109.1, β = 107.5, γ = 95.9°. The calculated values for V M and analysis of the self‐rotation and self‐Patterson functions suggest that the asymmetric unit in both crystal forms contains two dimers related by pseudo‐translational symmetry.