Crystallization and preliminary X-ray analysis of the haloalkane dehalogenase DatA from Agrobacterium tumefaciens C58.

Tomoko Mase, Hideya Yabuki, Masahiko Okai, Jun Ohtsuka, Fabiana Lica Imai, Yuji Nagata, Masaru Tanokura
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Abstract

Haloalkane dehalogenases are enzymes that catalyze the hydrolytic reaction of a wide variety of haloalkyl substrates to form the corresponding alcohol and hydrogen halide products. DatA from Agrobacterium tumefaciens C58 is a haloalkane dehalogenase that has a unique pair of halide-binding residues, asparagine (Asn43) and tyrosine (Tyr109), instead of the asparagine and tryptophan that are conserved in other members of the subfamily. DatA was expressed in Escherichia coli, purified and crystallized using the sitting-drop vapour-diffusion method with a reservoir solution consisting of 0.1 M CHES pH 8.6, 1.0 M potassium sodium tartrate, 0.2 M lithium sulfate, 0.01 M barium chloride. X-ray diffraction data were collected to 1.70 Å resolution. The space group of the crystal was determined as the primitive tetragonal space group P422, with unit-cell parameters a = b = 123.7, c = 88.1 Å. The crystal contained two molecules in the asymmetric unit.

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农杆菌 C58 中卤代烃脱卤酶 DatA 的结晶和初步 X 射线分析。
卤代烃脱卤酶是一种催化多种卤代烃底物水解反应生成相应的醇和卤化氢产物的酶。来自农杆菌 C58 的 DatA 是一种卤代烃脱卤酶,它有一对独特的卤化物结合残基,即天冬酰胺(Asn43)和酪氨酸(Tyr109),而不是亚家族其他成员中保留的天冬酰胺和色氨酸。DatA 在大肠杆菌中表达、纯化和结晶,采用坐滴蒸发扩散法,储液为 0.1 M CHES pH 8.6、1.0 M 酒石酸钾钠、0.2 M 硫酸锂和 0.01 M 氯化钡。X 射线衍射数据的分辨率为 1.70 Å。该晶体的空间群被确定为原始四方空间群 P422,单位晶胞参数为 a = b = 123.7,c = 88.1 Å。
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期刊介绍: Acta Crystallographica Section F is a rapid structural biology communications journal. Articles on any aspect of structural biology, including structures determined using high-throughput methods or from iterative studies such as those used in the pharmaceutical industry, are welcomed by the journal. The journal offers the option of open access, and all communications benefit from unlimited free use of colour illustrations and no page charges. Authors are encouraged to submit multimedia content for publication with their articles. Acta Cryst. F has a dedicated online tool called publBio that is designed to make the preparation and submission of articles easier for authors.
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