title: Molecular cloning, expression, purification and crystallographic analysis of zebrafish THEM2.
abstract: 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 C2, with unit-cell parameters a=77.1, b=74.4, c=96.6 Å, β=93.7°.
authors.affiliation: Laboratory of Non-coding RNA, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Beijing 100101, People's Republic of China.
authors.forename: Han
authors.initials: H
authors.lastname: Li
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
authors.forename: Shanshan
authors.initials: S
authors.lastname: Yu
authors.forename: Minze
authors.initials: M
authors.lastname: Jia
authors.forename: Weimin
authors.initials: W
authors.lastname: Gong
mesh_terms: D000214:Acyl Coenzyme A / Q000737:chemistry; D000818:Animals; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D010170:Palmitoyl-CoA Hydrolase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D014961:X-Ray Diffraction; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D000214:Acyl Coenzyme A; D029961:Zebrafish Proteins; D010170:Palmitoyl-CoA Hydrolase; C580996:acot13 protein, zebrafish