Open AccessCrystallization and preliminary X‐ray diffraction studies of a hyperthermophilic Rieske protein variant (SDX‐triple) with an engineered rubredoxin‐like mononuclear iron site
Author(s) -
Iwasaki Toshio,
Ohmori Daijiro,
Kounosu Asako,
Kumasaka Takashi
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106034476
Subject(s) - rubredoxin , pyrococcus furiosus , crystallography , crystallization , triclinic crystal system , chemistry , iron–sulfur cluster , x ray crystallography , ferredoxin , crystal structure , biochemistry , diffraction , enzyme , archaea , organic chemistry , physics , optics , gene
In place of the Rieske [2Fe–2S] cluster, an archetypal mononuclear iron site has rationally been designed into a hyperthermophilic archaeal Rieske [2Fe–2S] protein (sulredoxin) from Sulfolobus tokodaii by three residue replacements with reference to the Pyrococcus furiosus rubredoxin sequence. The resulting sulredoxin variant, SDX‐triple (H44I/A45C/H64C), has been purified and crystallized by the hanging‐drop vapour‐diffusion method using 65%( v / v ) 2‐methyl‐2,4‐pentanediol, 0.025 M citric acid and 0.075 M sodium acetate trihydrate pH 4.3. The crystals diffract to 1.63 Å resolution and belong to the triclinic space group P 1, with unit‐cell parameters a = 43.56, b = 76.54, c = 80.28 Å, α = 88.12, β = 78.82, γ = 73.46°. The asymmetric unit contains eight protein molecules.