Crystallization of the functional domain of human thrombopoietin using an antigen-binding fragment derived from neutralizing monoclonal antibody

Ryota Kuroki*, Masako Hirose, Yoichi Kato, Michael D. Feese, Taro Tamada, Hideki Shigematsu, Hiroshi Watarai, Yoshitake Maeda, Tomoyuki Tahara, Takashi Kato, Hiroshi Miyazaki

*この研究の対応する著者

研究成果: Article査読

9 被引用数 (Scopus)

抄録

Thrombopoietin (TPO) is a cytokine which primarily stimulates megakaryocytopoiesis and thrombopoiesis. The functional domain of TPO (TPO163) consisting of the N-terminal 163 amino acids was prepared and crystallized. Since the crystallization of TPO163 was unsuccessful using the standard screening methods, a Fab fragment derived from a neutralizing monoclonal antibody was used for crystallization. It was found that the TPO163-Fab complex crystallized reproducibly in 0.1 M potassium phosphate buffer pH 6.0 containing 20-25% polyethylene glycol 4000. Thin crystals (0.2 × 0.2 × 0.02 mm) grew in two space groups: P21, with unit-cell parameters a = 133.20, b = 46.71, c = 191.47 Å, β = 90.24°, and C2, with unit-cell parameters a = 131.71, b = 46.48, c = 184.63 Å, β = 90.42°. The results of a molecular-replacement analysis indicate that the Fab molecules interact with each other and provide a suitable interface for crystallization.

本文言語English
ページ(範囲)856-858
ページ数3
ジャーナルActa Crystallographica Section D: Biological Crystallography
58
5
DOI
出版ステータスPublished - 2002 1 1
外部発表はい

ASJC Scopus subject areas

  • 構造生物学

フィンガープリント

「Crystallization of the functional domain of human thrombopoietin using an antigen-binding fragment derived from neutralizing monoclonal antibody」の研究トピックを掘り下げます。これらがまとまってユニークなフィンガープリントを構成します。

引用スタイル