| Project/Area Number |
03670256
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | University of Tokyo (1992)
Kumamoto University (1991) |
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Principal Investigator |
YASUMICHI Hitoshi Inst. Med. Sci., Univ. Tokyo, Asst., 医科学研究所, 助手 (10222241)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGUCHI Naoto Inst. Mol. Embry. Genet., Kumamoto Univ. Med. Sch, Asst., 医学部, 助手 (00166620)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1992: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | Interleukin 5 / Interleukin 5 receptor / signal transduction / tyrosine phosphorylation / Interleukin 5 receptor alpha chain / Interleukin 5 receptor beta chain / インターロイキン5受容体α鎖 / インタ-ロイキン5 / インタ-ロイキン5受容体 / リン酸化蛋白 |
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| Research Abstract |
Murine interleukin 5 (IL-5) binds to its receptor with high and low affinity. It has been shown that the high affinity IL-5 receptor (IL-5R) is composed of at least two membrane protein subunits, p60 and p130, which are designated alpha chain and beta chain, respectively. The murine IL-5R alpha chain (IL-5Ralpha) was purified from IL-5 dependent early B cell line (T88-M) by using an immobilized monoclonal antibody against the murine IL-5Ralpha. The determined N-terminal sequence of the immunoaffinity-purified IL-5Ralpha corresponded precisely to the deduced primary sequence from cDNA encoding the IL-5Ralpha. Our data indicate that the signal peptide is N-terminal 17 aminoacids of the deduced sequence from the IL-5Ralpha cDNA. The high affinity IL-5R was reconstituted on an L-cells transfectant co-expressing IL-5Ralpha and AIC2B, which is the homologue of IL-3 receptor (AIC2A) and does not bind IL-5 its self. AIC2B is a component of the high affinity IL-5R, which is called beta chain. The rapid phosphorylation of a p60, which is not IL-5Ralpha, at serine residues and of p140, p92, p53, p48 and p45 at tyrosine residues were induced by the stimulation of T88-M cells with IL-5. Furthermore we investigated IL-5-induced tyrosine phosphorylation of IL-5Rbeta with an monoclonal antibody against the murine IL-5Rbeta. IL-5Rbeta appeared to be phosphorylated at tyrosine residues within 30 min after the stimulation with IL-5. The tyrosine-phosphorylated IL-5Rbeta disappeared 30 min after the stimulation. Interestingly, molecular weight of the tyrosine- phosphorylated IL-5Rbeta at 5 min after the stimulation was heavier than that at 1 min after the stimulation. Now we investigate association between IL-5Rbeta and other phosphorylated proteins with transfectants expressing mutant IL-5Ralpha and IL-5Rbeta.
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