Watanabe Aya, Hirata Kunio, Hagiwara Yoshinori, Yutani Yuko, Sugishima Masakazu, Yamamoto Masaki, Fukuyama Keiichi, Wada Kei
Department of Biological Sciences, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Mar 1;67(Pt 3):313-7. doi: 10.1107/S1744309110053431. Epub 2011 Feb 18.
Biliverdin reductase (BVR) catalyzes the conversion of biliverdin IX α to bilirubin IX α with concomitant oxidation of an NADH or NADPH cofactor. This enzyme also binds DNA and enhances the transcription of specific genes. Recombinant cyanobacterial BVR was overexpressed in Escherichia coli, purified and crystallized. A native data set was collected to 2.34 Å resolution on beamline BL38B1 at SPring-8. An SeMet data set was collected from a microcrystal (300×10×10 µm) on the RIKEN targeted protein beamline BL32XU and diffraction spots were obtained to 3.0 Å resolution. The native BVR crystal belonged to space group P2(1)2(1)2(1), with unit-cell parameters a=58.8, b=88.4, c=132.6 Å. Assuming that two molecules are present in the asymmetric unit, VM (the Matthews coefficient) was calculated to be 2.37 Å3 Da(-1) and the solvent content was estimated to be 48.1%. The structure of cyanobacterial BVR may provide insights into the mechanisms of its enzymatic and physiological functions.
胆绿素还原酶（BVR）催化胆绿素IXα转化为胆红素IXα，同时氧化NADH或NADPH辅因子。这种酶还能结合DNA并增强特定基因的转录。重组蓝藻BVR在大肠杆菌中过量表达、纯化并结晶。在SPring-8的BL38B1光束线上收集了分辨率为2.34 Å的天然数据集。从一个微晶（300×10×10 µm）在RIKEN靶向蛋白质光束线BL32XU上收集了硒代甲硫氨酸数据集，获得了分辨率为3.0 Å的衍射斑点。天然BVR晶体属于空间群P2(1)2(1)2(1)，晶胞参数a = 58.8、b = 88.4、c = 132.6 Å。假设不对称单位中有两个分子，计算出VM（马修斯系数）为2.37 Å3 Da(-1)，溶剂含量估计为48.1%。蓝藻BVR的结构可能为其酶促和生理功能机制提供见解。
