title: Complex assembly, crystallization and preliminary X-ray crystallographic analysis of the human Rod-Zwilch-ZW10 (RZZ) complex.
abstract: The spindle-assembly checkpoint (SAC) monitors kinetochore-microtubule attachment during mitosis. In metazoans, the three-subunit Rod-Zwilch-ZW10 (RZZ) complex is a crucial SAC component that interacts with additional SAC-activating and SAC-silencing components, including the Mad1-Mad2 complex and cytoplasmic dynein. The RZZ complex contains two copies of each subunit and has a predicted molecular mass of ∼800 kDa. Given the low abundance of the RZZ complex in natural sources, its recombinant reconstitution was attempted by co-expression of its subunits in insect cells. The RZZ complex was purified to homogeneity and subjected to systematic crystallization attempts. Initial crystals containing the entire RZZ complex were obtained using the sitting-drop method and were subjected to optimization to improve the diffraction resolution limit. The crystals belonged to space group P3₁ (No. 144) or P3₂ (No. 145), with unit-cell parameters a = b = 215.45, c = 458.7 Å, α = β = 90.0, γ = 120.0°.
authors: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Otto Hahn Strasse 11, 44227 Dortmund, Germany.
authors: Anika
authors: A
authors: Altenfeld
authors: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Otto Hahn Strasse 11, 44227 Dortmund, Germany.
authors: Sabine
authors: S
authors: Wohlgemuth
authors: Department `Genotoxic Stress and Cancer', Institut Curie, CNRS UMR 3348/INSERM U1005, Bâtiment 110, Centre Universitaire, 91405 Orsay CEDEX, France.
authors: Annemarie
authors: A
authors: Wehenkel
authors: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Otto Hahn Strasse 11, 44227 Dortmund, Germany.
authors: Ingrid R
authors: IR
authors: Vetter
authors: Department of Mechanistic Cell Biology, Max Planck Institute of Molecular Physiology, Otto Hahn Strasse 11, 44227 Dortmund, Germany.
authors: Andrea
authors: A
authors: Musacchio
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000235:genetics*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D007313:Insecta; D059566:M Phase Cell Cycle Checkpoints / Q000235:genetics*; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D008869:Microtubule-Associated Proteins; C488261:ZW10 protein, human
keywords: KMN network; Knl1; Mis12; Ndc80; RZZ complex; Rod; ZW10; Zwilch; cell division; kinetochore; mitosis; spindle-assembly checkpoint