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, ?=93.7°.

SUBMITTER: Li H 

PROVIDER: S-EPMC3509980 | biostudies-other | 2012 Dec

REPOSITORIES: biostudies-other