Crystallization of a 2:2 complex of granulocyte-colony stimulating factor (GCSF) with the ligand-binding region of the GCSF receptor.
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.
Honjo
Eijiro
E
Pharmaceutical Research Laboratory, Kirin Brewery Co. Ltd, Takasaki 370-1295, Japan.
Tamada
Taro
T
Maeda
Yoshitake
Y
Koshiba
Takumi
T
Matsukura
Yasuko
Y
Okamoto
Tomoyuki
T
Ishibashi
Matsujiro
M
Tokunaga
Masao
M
Kuroki
Ryota
R
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
D008024:Ligands; D046911:Macromolecular Substances; D016188:Receptors, Granulocyte Colony-Stimulating Factor; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor