Ohkuri Takatoshi, Takeda Chika, Yoshida Yuichiro, Izuhara Kenji, Imoto Taiji, Ueda Tadashi
Department of Immunology, Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
Protein Expr Purif. 2007 Nov;56(1):48-53. doi: 10.1016/j.pep.2007.05.008. Epub 2007 May 27.
Interleukin-13 receptor alpha2 (IL-13Ralpha2) 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-13Ralpha2 (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-13Ralpha2 as glycoprotein from P. pastoris. The yield of purified extracellular domain of human IL-13Ralpha2 was 2mg from 1L of culture. From CD analysis, the 2D structure of the purified IL-13Ralpha2 showed the typical beta-sheet. ELISA of the purified IL-13Ralpha2 detected the binding activity for human IL-13. Thus, it was found that the active extracellular domain of human IL-13Ralpha2 was expressed from P. pastoris.
白细胞介素-13受体α2（IL-13Rα2）以高亲和力结合IL-13，并作为诱饵受体在IL-13信号传导中发挥重要作用。我们在甲基营养型酵母毕赤酵母中表达了人IL-13Rα2（1-313）的细胞外结构域。通过PAS染色的SDS-PAGE分析和蛋白质印迹分析检测到毕赤酵母中作为糖蛋白的人IL-13Rα2细胞外结构域的产物。从1升培养物中纯化的人IL-13Rα2细胞外结构域的产量为2毫克。通过圆二色性分析，纯化的IL-13Rα2的二维结构显示出典型的β-折叠。纯化的IL-13Rα2的酶联免疫吸附测定检测到对人IL-13的结合活性。因此，发现人IL-13Rα2的活性细胞外结构域在毕赤酵母中表达。
