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Crystallization of a 2:2 complex of Granulocyte-Colony Stimulating Factor (GCSF) with the ligand-binding region of the GCSF receptor

顆粒球刺激因子(GCSF)受容体リガンド結合部位とGCSFの2:2複合体の結晶化

本庄 栄二郎; 玉田 太郎; 前田 宜丈*; 小柴 琢己*; 松倉 康子*; 岡本 智之*; 石橋 松二郎*; 徳永 正雄*; 黒木 良太

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

顆粒球刺激因子(GCSF)受容体は顆粒球前駆体の分化や増殖を調節する刺激を細胞内へ伝える。その受容体のリガンド結合部位とGCSFの2:2複合体の結晶化を行った。結晶は1.0Mギ酸ナトリウムを含む0.1M酢酸緩衝液(pH4.6)の条件で結晶化した。空間群は${it P}$4$$_{1}$$2$$_{1}$$2(もしくは${it P}$4$$_{3}$$2$$_{1}$$2)で、セル長は${it a}$=${it b}$=110.1$AA , {it c}$=331.8$AA $であった。しかしながら5$AA $以上の回折データが収集できなかったことから、受容体を陰イオン交換クロマトグラフィーで精製し、再度結晶化を試みた。その結果、3$AA $以上の回折データが収集可能な新たな晶形の結晶が得られた。その結晶の空間群は${it P}$3$$_{1}$$21(or its enantiomorph ${it P}$3$$_{2}$$21)で、セル長は${it a}$=${it b}$=134.8, ${it c}$=105.7$AA $であった。

Granulocyte colony-stimulating factor (GCSF) receptor receives signals for regulating the proliferation and differentiation of the precursor cells of granulocytes. The complex composed of two GCSFs and two GCSF receptors was crystallized. The crystal of the complex was grown in 1.0 M sodium formate and 0.1 M sodium acetate (pH4.6). It belongs to the space group ${it P}$4$$_{1}$$2$$_{1}$$2 (or its enantiomorph ${it P}$4$$_{3}$$2$$_{1}$$2) with unit cell dimensions of ${it a}$ = ${it b}$ = 110.1 ${AA}$, ${it c}$ = 331.8 ${AA}$. However, the diffraction data from the crystal beyond 5 ${AA}$ resolution could not be collected. Since the heterogeneity of GCSF receptor seems to interrupt growth of good quality crystals, the GCSF receptor was fractionated by achromatography. Crystals of GCSF/fractionated GCSF receptor complex were grown as a new crystal form in 0.2 M ammonium phosphate. The new crystal diffracts beyond 3.0 ${AA}$ resolution and belongs to space group ${it P}$3$$_{1}$$21 (or its enantiomorph ${it P}$3$$_{2}$$21) with unit-cell parameters ${it a}$ = ${it b}$ = 134.8, ${it c}$ = 105.7 ${AA}$.

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パーセンタイル:56.84

分野:Biochemical Research Methods

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