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

Research output: Contribution to journalArticlepeer-review

9 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)856-858
Number of pages3
JournalActa Crystallographica Section D: Biological Crystallography
Volume58
Issue number5
DOIs
Publication statusPublished - 2002 Jan 1
Externally publishedYes

ASJC Scopus subject areas

  • Structural Biology

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