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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

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Molecular cloning, expression, purification and crystallographic analysis of zebrafish THEM2.

Li Han H Gao Feng F Yu Shanshan S Jia Minze M Gong Weimin W

Acta crystallographica. Section F, Structural biology and crystallization communications 20121114 Pt 12

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 diffract ...[more]

PMID: 23192039

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