Open AccessCrystallization of a 2:2 complex of granulocyte‐colony stimulating factor (GCSF) with the ligand‐binding region of the GCSF receptor
Author(s) -
Honjo Eijiro,
Tamada Taro,
Maeda Yoshitake,
Koshiba Takumi,
Matsukura Yasuko,
Okamoto Tomoyuki,
Ishibashi Matsujiro,
Tokunaga Masao,
Kuroki Ryota
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105023080
Subject(s) - receptor , ligand (biochemistry) , crystallization , chemistry , crystal (programming language) , biophysics , materials science , crystallography , biochemistry , biology , organic chemistry , computer science , programming language
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 P 4 1 2 1 2 (or its enantiomorph P 4 3 2 1 2), with unit‐cell parameters a = b = 110.1, c = 331.8 Å. Unfortunately, this crystal form did not diffract beyond 5 Å 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 Å resolution and belonged to space group P 3 1 21 (or its enantiomorph P 3 2 21), with unit‐cell parameters a = b = 134.8, c = 105.7 Å.