Purification and characterization of the human interleukin-18 receptor
Purification and characterization of the human interleukin-18 receptor
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DOI：
10.1074/jbc.272.41.25737
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发表时间：
1997-10-10
影响因子：
4.8
通讯作者：
Kurimoto, M
中科院分区：
文献类型：
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作者：
Torigoe, K;Ushio, S;Kurimoto, M
白细胞介素18(IL-18)被认为是一种诱导产生干扰素-γ和增强NK细胞杀伤活性的分子。本文报道了人IL-18受体(hIL-18R)的纯化和鉴定。根据结合实验的结果，我们选择Hodgkin病细胞系L428作为hIL-18R表达最强的细胞系。这种结合可被IL-18抑制，但不被IL-1β抑制。I-125-IL-18与L428细胞结合的解离常数(K-d)约为18.5 nm，结合位点数为18,000个/细胞。用L428细胞免疫小鼠并克隆后，获得了一株抗hIL-18R的单抗(mAb117-10C)。3-[(3-胆酰胺丙基)二甲基氨基]-1-丙磺酸(CHAPS)提取的L428细胞经小麦胚凝集素-Sepharose4B柱层析和mAb117-10C-Sepharose柱层析纯化得到hIL-18R。HIL-18R的内部氨基酸序列都与人IL-1受体相关蛋白(IL-1Rrp)的氨基酸序列一致，但其配体尚不清楚。当IL-1Rrp在COS-1细胞中表达时，可使细胞具有与IL-18结合的特性和信号转导能力。根据这些结果，我们得出结论，IL-18受体的一个功能成分是IL-1Rrp。
Interleukin (IL)-18 was identified as a molecule that induces IFN-gamma production and enhances NK cell cytotoxicity. In this paper, we report upon the purification and characterization of human IL-18 receptor (hIL-18R). We selected the Hodgkin's disease cell line, L428, as the most strongly hIL-18R-expressing cell line based on the results of binding assays. This binding was inhibited by IL-18 but not by IL-1 beta. The dissociation constant (K-d) of I-125-IL-18 binding to L428 cells was about 18.5 nM, with 18,000 binding sites/cell. After immunizing mice with L428 cells and cloning, a single monoclonal antibody (mAb) against hIL-18R was obtained (mAb 117-10C). Sequentially, hIL-18R was purified from 3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonic acid (CHAPS)-extracted L428 cells by wheat germ lectin-Sepharose 4B chromatography and mAb 117-10C-Sepharose chromatography. The internal amino acid sequences of hIL-18R all matched those of human IL-1 receptor-related protein (IL-1Rrp), the ligand of which was unknown to date. When expressed in COS-1 cells, the cDNA of IL-1Rrp conferred IL-18 binding properties on the cells and the capacity for signal transduction. From these results, we conclude that a functional IL-18 receptor component is IL-1Rrp.