Open AccessMolecular cloning, expression, purification and crystallographic analysis of zebrafish THEM2
Author(s) -
Li Han,
Gao Feng,
Yu Shanshan,
Jia Minze,
Gong Weimin
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112043813
Subject(s) - thioesterase , escherichia coli , thioester , crystallography , chemistry , size exclusion chromatography , cloning (programming) , monoclinic crystal system , affinity chromatography , biochemistry , crystal structure , enzyme , biosynthesis , gene , computer science , programming language
Thioesterase superfamily member 2 (THEM2) is essential for cell proliferation of mammalian cells. It belongs to the hotdog‐fold thioesterase superfamily and catalyzes the hydrolysis of the thioester bonds of acyl‐CoA in vitro . In this study, THEM2 protein from zebrafish (fTHEM2) was expressed in Escherichia coli and purified by Ni‐affinity and gel‐filtration chromatography. fTHEM2 crystals were obtained using the sitting‐drop vapour‐diffusion method with PEG 10 000 as precipitant. X‐ray diffraction data were collected to 1.80 Å resolution using a synchrotron‐radiation source. The crystals belonged to the monoclinic space group C 2, with unit‐cell parameters a = 77.1, b = 74.4, c = 96.6 Å, β = 93.7°.