Expression of human IL-13 receptor α2 extracellular domain in Pichia pastoris

Takatoshi Ohkuri; Chika Takeda; Yuichiro Yoshida; Kenji Izuhara; Taiji Imoto; Tadashi Ueda

doi:10.1016/j.pep.2007.05.008

Expression of human IL-13 receptor α2 extracellular domain in Pichia pastoris

Takatoshi Ohkuri, Chika Takeda, Yuichiro Yoshida, Kenji Izuhara, Taiji Imoto, Tadashi Ueda

Pharmaceutical Health Care and Sciences

Research output: Contribution to journal › Article › peer-review

2 Citations (Scopus)

Abstract

Interleukin-13 receptor α2 (IL-13Rα2) binds IL-13 with high affinity and plays an important role in IL-13 signaling as a decoy receptor. We expressed the extracellular domain of human IL-13Rα2 (1-313) in methylotrophic yeast Pichia pastoris. SDS-PAGE analysis by PAS staining and Western blot analysis detected the product of the extracellular domain of human IL-13Rα2 as glycoprotein from P. pastoris. The yield of purified extracellular domain of human IL-13Rα2 was 2 mg from 1 L of culture. From CD analysis, the 2D structure of the purified IL-13Rα2 showed the typical β-sheet. ELISA of the purified IL-13Rα2 detected the binding activity for human IL-13. Thus, it was found that the active extracellular domain of human IL-13Rα2 was expressed from P. pastoris.

Original language	English
Pages (from-to)	48-53
Number of pages	6
Journal	Protein Expression and Purification
Volume	56
Issue number	1
DOIs	https://doi.org/10.1016/j.pep.2007.05.008
Publication status	Published - Nov 1 2007

All Science Journal Classification (ASJC) codes

Biotechnology

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10.1016/j.pep.2007.05.008

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title = "Expression of human IL-13 receptor α2 extracellular domain in Pichia pastoris",

abstract = "Interleukin-13 receptor α2 (IL-13Rα2) binds IL-13 with high affinity and plays an important role in IL-13 signaling as a decoy receptor. We expressed the extracellular domain of human IL-13Rα2 (1-313) in methylotrophic yeast Pichia pastoris. SDS-PAGE analysis by PAS staining and Western blot analysis detected the product of the extracellular domain of human IL-13Rα2 as glycoprotein from P. pastoris. The yield of purified extracellular domain of human IL-13Rα2 was 2 mg from 1 L of culture. From CD analysis, the 2D structure of the purified IL-13Rα2 showed the typical β-sheet. ELISA of the purified IL-13Rα2 detected the binding activity for human IL-13. Thus, it was found that the active extracellular domain of human IL-13Rα2 was expressed from P. pastoris.",

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AU - Ohkuri, Takatoshi

AU - Takeda, Chika

AU - Yoshida, Yuichiro

AU - Izuhara, Kenji

AU - Imoto, Taiji

AU - Ueda, Tadashi

PY - 2007/11/1

Y1 - 2007/11/1

N2 - Interleukin-13 receptor α2 (IL-13Rα2) binds IL-13 with high affinity and plays an important role in IL-13 signaling as a decoy receptor. We expressed the extracellular domain of human IL-13Rα2 (1-313) in methylotrophic yeast Pichia pastoris. SDS-PAGE analysis by PAS staining and Western blot analysis detected the product of the extracellular domain of human IL-13Rα2 as glycoprotein from P. pastoris. The yield of purified extracellular domain of human IL-13Rα2 was 2 mg from 1 L of culture. From CD analysis, the 2D structure of the purified IL-13Rα2 showed the typical β-sheet. ELISA of the purified IL-13Rα2 detected the binding activity for human IL-13. Thus, it was found that the active extracellular domain of human IL-13Rα2 was expressed from P. pastoris.

AB - Interleukin-13 receptor α2 (IL-13Rα2) binds IL-13 with high affinity and plays an important role in IL-13 signaling as a decoy receptor. We expressed the extracellular domain of human IL-13Rα2 (1-313) in methylotrophic yeast Pichia pastoris. SDS-PAGE analysis by PAS staining and Western blot analysis detected the product of the extracellular domain of human IL-13Rα2 as glycoprotein from P. pastoris. The yield of purified extracellular domain of human IL-13Rα2 was 2 mg from 1 L of culture. From CD analysis, the 2D structure of the purified IL-13Rα2 showed the typical β-sheet. ELISA of the purified IL-13Rα2 detected the binding activity for human IL-13. Thus, it was found that the active extracellular domain of human IL-13Rα2 was expressed from P. pastoris.

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Expression of human IL-13 receptor α2 extracellular domain in Pichia pastoris

Abstract

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