title: Crystallization of a 2:2 complex of granulocyte-colony stimulating factor (GCSF) with the ligand-binding region of the GCSF receptor.
abstract: The granulocyte-colony stimulating factor (GCSF) receptor receives signals for regulating the maturation, proliferation and differentiation of the precursor cells of neutrophilic granulocytes. The signalling complex composed of two GCSFs (GCSF, 19 kDa) and two GCSF receptors (GCSFR, 34 kDa) consisting of an Ig-like domain and a cytokine-receptor homologous (CRH) domain was crystallized. A crystal of the complex was grown in 1.0 M sodium formate and 0.1 M sodium acetate pH 4.6 and belongs to space group P4(1)2(1)2 (or its enantiomorph P4(3)2(1)2), with unit-cell parameters a = b = 110.1, c = 331.8 A. Unfortunately, this crystal form did not diffract beyond 5 A resolution. Since the heterogeneity of GCSF receptor appeared to prevent the growth of good-quality crystals, the GCSF receptor was fractionated by anion-exchange chromatography. Crystals of the GCSF-fractionated GCSF receptor complex were grown as a new crystal form in 0.2 M ammonium phosphate. This new crystal form diffracted to beyond 3.0 A resolution and belonged to space group P3(1)21 (or its enantiomorph P3(2)21), with unit-cell parameters a = b = 134.8, c = 105.7 A.
authors.affiliation: Pharmaceutical Research Laboratory, Kirin Brewery Co. Ltd, Takasaki 370-1295, Japan.
authors.forename: Eijiro
authors.initials: E
authors.lastname: Honjo
authors.forename: Taro
authors.initials: T
authors.lastname: Tamada
authors.forename: Yoshitake
authors.initials: Y
authors.lastname: Maeda
authors.forename: Takumi
authors.initials: T
authors.lastname: Koshiba
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Matsukura
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Okamoto
authors.forename: Matsujiro
authors.initials: M
authors.lastname: Ishibashi
authors.forename: Masao
authors.initials: M
authors.lastname: Tokunaga
authors.forename: Ryota
authors.initials: R
authors.lastname: Kuroki
mesh_terms: D002852:Chromatography, Ion Exchange; D003460:Crystallization; D004591:Electrophoresis, Polyacrylamide Gel; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D046911:Macromolecular Substances / Q000737:chemistry*; D016188:Receptors, Granulocyte Colony-Stimulating Factor / Q000737:chemistry* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D008024:Ligands; D046911:Macromolecular Substances; D016188:Receptors, Granulocyte Colony-Stimulating Factor; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor