Crystallization of a 2:2 complex of granulocyte-colony stimulating factor (GCSF) with the ligand-binding region of the GCSF receptor

Eijiro Honjo, Taro Tamada, Yoshitake Maeda, Takumi Koshiba, Yasuko Matsukura, Tomoyuki Okamoto, Matsujiro Ishibashi, Masao Tokunaga, Ryota Kuroki

Research output: Contribution to journalArticle

8 Citations (Scopus)

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 P41212 (or its enantiomorph P43212), 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 P3121 (or its enantiomorph P3221), with unit-cell parameters a = b = 134.8, c = 105.7 Å.

Original languageEnglish
Pages (from-to)788-790
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume61
Issue number8
DOIs
Publication statusPublished - Aug 1 2005

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Granulocyte Colony-Stimulating Factor Receptors
Granulocyte Colony-Stimulating Factor
Crystallization
crystallization
Ligands
Crystals
ligands
formic acid
crystals
Sodium Acetate
Granulocyte Precursor Cells
Cytokine Receptors
cells
sodium
ammonium phosphates
Anions
Chromatography
formates
chromatography
acetates

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

Cite this

Crystallization of a 2:2 complex of granulocyte-colony stimulating factor (GCSF) with the ligand-binding region of the GCSF receptor. / Honjo, Eijiro; Tamada, Taro; Maeda, Yoshitake; Koshiba, Takumi; Matsukura, Yasuko; Okamoto, Tomoyuki; Ishibashi, Matsujiro; Tokunaga, Masao; Kuroki, Ryota.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 61, No. 8, 01.08.2005, p. 788-790.

Research output: Contribution to journalArticle

Honjo, Eijiro ; Tamada, Taro ; Maeda, Yoshitake ; Koshiba, Takumi ; Matsukura, Yasuko ; Okamoto, Tomoyuki ; Ishibashi, Matsujiro ; Tokunaga, Masao ; Kuroki, Ryota. / Crystallization of a 2:2 complex of granulocyte-colony stimulating factor (GCSF) with the ligand-binding region of the GCSF receptor. In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications. 2005 ; Vol. 61, No. 8. pp. 788-790.
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