Purification and Characterization of the Human Interleukin-18 Receptor
| Autor: | Toshio Kunikata, Hirotada Kojima, Shimpei Ushio, Takanori Okura, Masashi Kurimoto, Mitsukiyo Fujii, Osamu Sanou, Hakuo Ikegami, Susumu Kobayashi, Tadatoshi Murakami, Kakuji Torigoe, Madoka Taniai, Tsunetaka Ohta, Masao Ikeda |
|---|---|
| Rok vydání: | 1997 |
| Předmět: |
medicine.drug_class
Molecular Sequence Data Biology Monoclonal antibody Biochemistry Mice Tumor Cells Cultured medicine Animals Humans Amino Acid Sequence Binding site Receptor Molecular Biology Mice Inbred BALB C Receptors Interleukin-18 Cell Membrane Interleukin-18 NF-kappa B Antibodies Monoclonal Interleukin Receptors Interleukin Cell Biology Ligand (biochemistry) Hodgkin Disease Molecular biology Interleukin-18 receptor Kinetics Cell culture COS Cells Cytokines Signal transduction Interleukin-18 Receptor alpha Subunit Interleukin-1 |
| Zdroj: | Journal of Biological Chemistry. 272:25737-25742 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.272.41.25737 |
| Popis: | Interleukin (IL)-18 was identified as a molecule that induces IFN-γ 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-1β. The dissociation constant (K d) of125I-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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|