title: Expression, purification and preliminary crystallographic analysis of the cryptic polo-box domain of Caenorhabditis elegans ZYG-1.
abstract: 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.
authors.affiliation: Department of Medical Biochemistry, Max F. Perutz Laboratories, Medical University of Vienna, Dr Bohr-Gasse 9, 1030 Vienna, Austria.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Shimanovskaya
authors.affiliation: Department of Medical Biochemistry, Max F. Perutz Laboratories, Medical University of Vienna, Dr Bohr-Gasse 9, 1030 Vienna, Austria.
authors.forename: Gang
authors.initials: G
authors.lastname: Dong
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D029742:Caenorhabditis elegans Proteins; D011494:Protein Kinases; C429649:zyg-1 protein, C elegans
keywords: Plk4; ZYG-1; centriole; cryptic polo-box domain; polo-like kinase