title: Purification, crystallization and preliminary crystallographic analysis of very-long-chain acyl-CoA dehydrogenase from Caenorhabditis elegans.
abstract: Acyl-CoA dehydrogenase [acyl-CoA:(acceptor) 2,3-oxidoreductase; EC 1.3.99.3] catalyzes the first reaction step in mitochondrial fatty-acid beta-oxidation. Here, the very-long-chain acyl-CoA dehydrogenase from Caenorhabditis elegans (cVLCAD) has been cloned and overexpressed in Escherichia coli strain BL21 (DE3). Interestingly, unlike other very-long-chain acyl-CoA dehydrogenases, cVLCAD was found to form a tetramer by size-exclusion chromatography coupled with in-line static light-scattering, refractive-index and ultraviolet measurements. Purified cVLCAD (12 mg ml(-1)) was successfully crystallized by the hanging-drop vapour-diffusion method under conditions containing 100 mM Tris-HCl pH 8.0, 150 mM sodium chloride, 200 mM magnesium formate and 13% PEG 3350. The crystal has a tetragonal form and a complete diffraction data set was collected and processed to 1.8 A resolution. The crystal belonged to space group C2, with unit-cell parameters a = 138.6, b = 116.7, c = 115.3 A, alpha = gamma = 90.0, beta = 124.0 degrees . A self-rotation function indicated the existence of one noncrystallographic twofold axis. A preliminary molecular-replacement solution further confirmed the presence of two molecules in one asymmetric unit, which yields a Matthews coefficient V(M) of 2.76 A(3) Da(-1) and a solvent content of 55%.
authors.affiliation: National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, People's Republic of China.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Li
authors.forename: Yujia
authors.initials: Y
authors.lastname: Zhai
authors.forename: Junnan
authors.initials: J
authors.lastname: Fang
authors.forename: Qiangjun
authors.initials: Q
authors.lastname: Zhou
authors.forename: Yunqi
authors.initials: Y
authors.lastname: Geng
authors.forename: Fei
authors.initials: F
authors.lastname: Sun
mesh_terms: D044942:Acyl-CoA Dehydrogenase, Long-Chain / Q000737:chemistry* / Q000302:isolation & purification; D000818:Animals; D017173:Caenorhabditis elegans / Q000201:enzymology*; D003460:Crystallization; D018360:Crystallography, X-Ray; D008970:Molecular Weight
chemical_list: D044942:Acyl-CoA Dehydrogenase, Long-Chain
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