| S(yp) Y279和Y304介导了BcrAbl与Grb2等蛋白的结合 |
| |
| 引用本文： | 朱奎,刘小萍,叶芳耘.S(yp) Y279和Y304介导了BcrAbl与Grb2等蛋白的结合[J].生物化学与生物物理学报,1999(4). |
| |
| 作者姓名： | 朱奎 刘小萍 叶芳耘 |
| |
| 作者单位： | 第二军医大学长海医院实验诊断科/中心实验室(朱奎，叶芳耘)，东方肝胆外科研究所肿瘤免疫与基因治疗中心!上海200433(刘小萍) |
| |
| 摘 要： | 利用体外定点突变技术获得Ｓｙｐ Ｙ２７９Ｆ、Ｙ３０４Ｆ和Ｙ５４６Ｆ突变的ｃＤＮＡ， 将这些突变体和野生型Ｓｙｐ 分别构建入ｐＸＭ 真核表达载体， 转入Ｋ５６２ 细胞。经Ｗｅｓｔｅｒｎ 印迹证明， 各转染Ｋ５６２ 细胞中都有Ｓｙｐ 蛋白的表达。免疫沉淀与免疫印迹结果发现ＷＴ、Ｙ２７９Ｆ、Ｙ３０４Ｆ和Ｙ５４６Ｆ等４ 种Ｓｙｐ 在胞内均能直接与ＢｃｒＡｂｌ 结合。体外结合实验结果表明Ｙ３０４Ｆ突变导致了Ｓｙｐ 不能与Ｓｈｃ 结合，Ｙ２７９Ｆ突变则导致了Ｓｙｐ 不能与Ｇｒｂ２ 结合。结论是： 作为“接头蛋白”，Ｓｙｐ 可以介导ＢｃｒＡｂｌ 与Ｓｈｃ 和Ｇｒｂ２ 之间的结合；Ｇｒｂ２ 结合在Ｓｙｐ 的Ｙ２７９ 上，Ｓｈｃ 则结合在Ｓｙｐ的Ｙ３０４ 上
|
| 关 键 词： | Bcr Abl激酶 Syp 信号传导 |
Syp Y279, Y304 Can Mediate the Binding of Bcr Abl to Grb2 and Other Proteins |
| |
| ZHU Kui ,LIU Xiao Ping ,ZENG Grant and YE Fang Yun.Syp Y279, Y304 Can Mediate the Binding of Bcr Abl to Grb2 and Other Proteins[J].Acta Biochimica et Biophysica Sinica,1999(4). |
| |
| Authors: | ZHU Kui LIU Xiao Ping ZENG Grant and YE Fang Yun |
| |
| Institution: | ZHU Kui 1*,LIU Xiao Ping 2,ZENG Grant 3 and YE Fang Yun 1 |
| |
| Abstract: | In this study we obtained 3 mutants of Syp cDNA Y279F, Y304F and Y546F by using the method of site directed mutation in vitro. We then inserted them into the pXM mammalian expression vector, and transferred the plasmids into K562 cells. Western Blot has found WT/ 3MT Syp are expressed in the K562 cells. Immunoprecipitation and Immunoblot have found that WT, Y279F, Y304F and Y546F Syp can bind directly to the Bcr Abl in vivo. However, we found that mutation of Syp Y279F can block the binding of Grb2 SH2 to Syp; mutation of Syp Y304F can block the binding of Shc SH2 to Syp in vitro. As a adaptor, Syp can mediate binding of Bcr Abl to Shc and Grb2. Grb2 bind to Syp pY279, Shc bind to Syp pY304 in vitro. |
| |
| Keywords: | Bcr Abl tyrosine kinase Syp protein signal transduction |
| 本文献已被 CNKI 等数据库收录！ |
