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Molecular analysis of signal transduction through IL-5 receptor

IL-5 受体信号转导的分子分析

基本信息

批准号：
03670256
负责人：
YASUMICHI Hitoshi
金额：
$ 1.28万
依托单位：
University of Tokyo (1992)Kumamoto University (1991)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1992
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03670256/
关键词：
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

项目摘要

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.

小鼠白细胞介素5（IL-5）以高和低亲和力与其受体结合。高亲和力IL-5受体（IL-5 R）由至少两个膜蛋白亚基p60和p130组成，分别命名为α链和β链。通过使用针对鼠IL-5 R α的固定化单克隆抗体，从IL-5依赖性早期B细胞系（T88-M）纯化鼠IL-5 R α链（IL-5 R α）。免疫亲和纯化的IL-5 R α的确定的N-末端序列精确地对应于从编码IL-5 R α的cDNA推导的一级序列。我们的数据表明，信号肽是从IL-5 R α cDNA推导的序列的N-末端17个氨基酸。高亲和力IL-5 R在共表达IL-5 R α和AIC 2B的L-细胞转染子上重建，AIC 2B是IL-3受体（AIC 2A）的同源物并且不结合IL-5本身。AIC 2B是高亲和力IL-5 R的组成部分，称为β链。通过用IL-5刺激T88-M细胞，诱导p60（不是IL-5 R α）在丝氨酸残基和p140、p92、p53、p48和p45在酪氨酸残基的快速磷酸化。此外，我们研究了IL-5诱导的酪氨酸磷酸化的IL-5 R β与单克隆抗体对鼠IL-5 R β。IL-5 R β在用IL-5刺激后30分钟内似乎在酪氨酸残基处磷酸化。酪氨酸磷酸化的IL-5 R β在刺激后30 min消失。有趣的是，刺激后5 min酪氨酸磷酸化的IL-5 R β的分子量比刺激后1 min的分子量大。现在，我们研究IL-5 R β和其他磷酸化蛋白质与表达突变IL-5 R α和IL-5 R β的转染子之间的关联。