Shimanovskaya Ekaterina, Dong Gang
Department of Medical Biochemistry, Max F. Perutz Laboratories, Medical University of Vienna, Dr Bohr-Gasse 9, 1030 Vienna, Austria.
Acta Crystallogr F Struct Biol Commun. 2014 Oct;70(Pt 10):1346-50. doi: 10.1107/S2053230X14016094. Epub 2014 Sep 25.
ZYG-1 is a polo-like kinase essential for centriole assembly in Caenorhabditis elegans. The targeting of ZYG-1 to nascent centrioles is via its central cryptic polo-box (CPB) domain. To shed light on the molecular basis of ZYG-1 recruitment, it is necessary to obtain structural knowledge of the ZYG-1 CPB. Here, the expression, purification and preliminary crystallographic analysis of the ZYG-1 CPB are reported. The protein was overexpressed in Escherichia coli strain BL21 (DE3), purified by multi-step chromatography and crystallized using the vapour-diffusion method. Crystals of the wild-type protein exhibited an order-disorder pathology, which was solved by reductive lysine methylation. A complete anomalous data set was collected to 2.54 Å resolution at the Se K edge (λ = 0.9792 Å). The crystal belonged to space group P2, with unit-cell parameters a = 53.3, b = 60.09, c = 87.51 Å, β = 93.31°. There were two molecules in the asymmetric unit.
ZYG-1是一种在秀丽隐杆线虫中对中心粒组装至关重要的类polo激酶。ZYG-1靶向新生中心粒是通过其中心的隐蔽polo盒（CPB）结构域实现的。为了阐明ZYG-1招募的分子基础，有必要获得ZYG-1 CPB的结构知识。在此，报道了ZYG-1 CPB的表达、纯化及初步晶体学分析。该蛋白在大肠杆菌BL21（DE3）菌株中过表达，通过多步层析纯化，并采用气相扩散法结晶。野生型蛋白的晶体表现出有序-无序病理现象，通过还原赖氨酸甲基化得以解决。在硒K边（λ = 0.9792 Å）收集了完整的反常数据集，分辨率达到2.54 Å。晶体属于空间群P2，晶胞参数为a = 53.3、b = 60.09、c = 87.51 Å，β = 93.31°。不对称单元中有两个分子。
