Mase Tomoko, Yabuki Hideya, Okai Masahiko, Ohtsuka Jun, Imai Fabiana Lica, Nagata Yuji, Tanokura Masaru
Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Jun 1;68(Pt 6):652-4. doi: 10.1107/S1744309112013942. Epub 2012 May 23.
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.
卤代烷脱卤酶是一类催化多种卤代烷基底物发生水解反应，生成相应醇类和卤化氢产物的酶。根癌土壤杆菌C58的DatA是一种卤代烷脱卤酶，它具有一对独特的卤化物结合残基，即天冬酰胺（Asn43）和酪氨酸（Tyr109），而不是该亚家族其他成员中保守的天冬酰胺和色氨酸。DatA在大肠杆菌中表达，通过坐滴气相扩散法进行纯化和结晶，所用储液由0.1 M CHES（pH 8.6）、1.0 M酒石酸钾钠、0.2 M硫酸锂、0.01 M氯化钡组成。收集到分辨率为1.70 Å的X射线衍射数据。晶体的空间群被确定为原始四方空间群P422，晶胞参数a = b = 123.7，c = 88.1 Å。不对称单元中包含两个分子。
