Open AccessExpression, purification, crystallization and preliminary X‐ray diffraction analysis of Arabidopsis thaliana Deg8
Author(s) -
Shan Xiaoyue,
Sun Wei,
Fan Haitian,
Jia Minze,
Gao Feng,
Gong Weimin
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112048774
Subject(s) - arabidopsis thaliana , crystallization , crystallography , x ray crystallography , escherichia coli , trimer , chemistry , solvent , biophysics , biochemistry , diffraction , biology , mutant , dimer , physics , organic chemistry , gene , optics
Arabidopsis thaliana Deg8, an ATP‐independent serine endopeptidase, is involved in the repair of photosystem II (PSII), specifically the degradation of the photo‐damaged PSII reaction centre D1 protein. To understand the molecular mechanism underlying the participation of Deg8 in the degradation of the photo‐damaged D1 protein, the structure of Deg8 is needed. Until recently, however, no structure of Deg8 had been solved. In this study, Deg8 from A. thaliana was cloned, overexpressed and purified in Escherichia coli . Crystallization was performed at 277 K using tribasic sodium citrate as the precipitant and the crystals diffracted to 2.0 Å resolution, belonging to space group C 2 with unit‐cell parameters a = 129.5, b = 124.2, c = 93.3 Å, α = γ = 90, β = 132.4°. Assuming one trimer in the asymmetric unit, the Matthews coefficient and the solvent content were calculated to be 2.35 Å 3 Da −1 and 47.6%, respectively.