Molecular cloning of an insulin receptor tyrosine-kinase substrate-1 (IRS-1) associated molecule

本课题取得了以下成果：1.制备了一系列抗人IRS-1的单克隆抗体.以单克隆抗体和体外酪氨酸磷酸化的重组IRS-1为探针，从HeLa细胞cDNA文库中分离IRS-1相关蛋白的候选cDNA.对其中一个候选基因HH 109进行了分析，发现它是一个新基因.在IRS-1基因缺陷小鼠中，检测到额外的胰岛素应答酪氨酸磷酸化蛋白（pp 190）。我们发现pp 190被抗IRS-1单抗6G5.5识别。为了分离pp 190的cDNA，我们以单克隆抗体6 G5为探针，对表达文库进行了筛选。我们发现MoAb，6 G5识别IRS-2，这是与pp190.7相同的分子。HH 109与IRS-2之间没有序列相似性。

The following resalts were obtained in the project :1. A number of monoclonal antibodies (MoAbs) against human IRS-1 were produced.2. Using both MoAb and the recombinant IRS-1 tyrosine-phosphoryrated in vitro as probes candidate cDNAs of IRS-1 associated proteines were isolated from HeLa cell cDNA library.3. One of candidated cDNAs, named HH109 has been analyzed and was found to be a novel gene.4. In the IRS-1 gene deficient mice an additional insulin responsed tyrosine-phosphorylated protein (pp190) was detected. We showed that pp190 was recognized by anti IRS-1 MoAb, 6G5.5. To isolate the cDNA of pp190 we srceened the expression library with the MoAb, 6G5 as a probe. And we showed that MoAb, 6G5 recognized IRS-2 which was identical molecule to pp190.7. There was no sequence similarity between HH109 and IRS-2.