Open AccessCrystallization and preliminary X‐ray diffraction studies of the prototypal homologue of mitoNEET ( Tth ‐NEET0026) from the extreme thermophile Thermus thermophilus HB8
Author(s) -
Kounosu Asako,
Iwasaki Toshio,
Baba Seiki,
HayashiIwasaki Yoko,
Oshima Tairo,
Kumasaka Takashi
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108035975
Subject(s) - thermus thermophilus , thermophile , crystallization , crystallography , materials science , x ray , x ray crystallography , diffraction , chemistry , physics , optics , thermodynamics , gene , biochemistry , escherichia coli , enzyme
MitoNEET (a mammalian mitochondrial outer membrane protein) is a potential pharmacological and clinical target of the insulin‐sensitizer pioglitazone. The thermophilic homologue of mitoNEET (TTHA0026) from Thermus thermophilus HB8 has been heterologously overproduced in Escherichia coli and purified as a water‐soluble prototypal protein containing the mitoNEET‐like [2Fe–2S] cluster. The resultant recombinant protein, named Tth ‐NEET0026, has been crystallized in its oxidized form by the hanging‐drop vapour‐diffusion method using 17%( w / v ) polyethylene glycol 4000, 8.5%( v / v ) 2‐propanol, 15%( v / v ) glycerol and 0.085 M HEPES–NaOH pH 7.2. The dark reddish crystals diffracted to 1.80 Å resolution and belonged to the tetragonal space group P 4 3 2 1 2, with unit‐cell parameters a = 45.51, c = 84.26 Å. The asymmetric unit contains one protein molecule.