Open AccessGPX3 from Arabidopsis thaliana : cloning, expression, purification, crystallization and preliminary X‐ray analysis
Author(s) -
Li Kun,
Yang Qingzhan,
Wang Wei,
Zhao Xiaoliang,
Lou Zhiyong
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113025566
Subject(s) - gpx3 , biology , arabidopsis thaliana , cloning (programming) , gene , microbiology and biotechnology , escherichia coli , biochemistry , chemistry , glutathione , glutathione peroxidase , enzyme , mutant , computer science , programming language
The Arabidopsis thaliana glutathione peroxidase 3 (GPX3) gene encodes a glutathione peroxidase with roles in H 2 O 2 homeostasis and signalling. The GPX3 gene sequence was cloned into pGEX‐6P1 and overexpressed in Escherichia coli . The GPX3 protein was purified to homogeneity in two chromatographic steps. Various lengths of the GPX3 sequence were used to obtain proteins that yielded crystals using vapour‐diffusion techniques, but only GPX3ΔN36 (lacking 36 amino acids from the N‐terminus) showed a good diffraction pattern. Its crystals diffracted to 2.8 Å resolution and belonged to space group P 6 5 , with unit‐cell parameters a = b = 98.241, c = 42.057 Å.