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c-Mpl信号转导相关蛋白的结合域确定

Identification of the Binding Domain of Signal Transduction Related Proteins on c-Mpl

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【作者】 杨立敏冯丽冰黄莉周伟国万敏李昌本

【Author】 YANG Limin, FENG Libing, HUANG Li, ZHOU Weiguo, WAN Min, LI Changben(Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China)

【机构】 复旦大学生命科学学院遗传学研究所复旦大学生命科学学院遗传学研究所 上海 200433上海 200433上海 200433

【摘要】 血小板生成素(thrombopoietin,TPO)是调节血小板生成最主要的细胞因子,它的生物学效应由其受体c Mpl介导.为了确定SGK,14 3 3,Siva,Tom1,Cop9 S3和c MplBP等6种蛋白在c Mpl上的结合区域,构建c Mpl膜内部分系列缺失片段及点突变,用酵母双杂合的方法检测这些片段与上述6种蛋白的结合情况,发现Box1是c Mpl与这6种蛋白结合的主要区域.Box1两侧的序列及C端区域对c Mpl与这6种蛋白的结合也有帮助.将c Mpl膜内部分的第531位Ser(位于Box1内)突变为Ala和Val后,c Mpl与SGK,14 3 3,Tom1和Cop9 S3的结合能力明显下降,提示Ser531可能对c Mpl与这4种蛋白的结合起重要作用.

【Abstract】 cMpl is the receptor of thrombopoietin, the critical regulator of platelet production. Proteins SGK, 1433, Siva, Tom1, Cop9S3 and cMplBP have recently been proved to interact with cMpl. With the intent to identify the binding domains of the 6 proteins on cMpl, serial deletions and point mutations of the cytotail of cMpl have been generated and their interaction with the proteins mentioned above have been examined by yeast two hybrid system. The results indicate that Box1 (a highly conserved region within the cytokine receptor superfamily) is the critical domain for binding. Considering the similar role Box1 plays in other cytokine receptors, the results suggest that Box1 is probably a widely used proteinbinding domain in cytokine receptors. Amino acids adjacent to Box1 and the Cterminal region also contribute to the interaction. Point mutations of Ser531 have negative effects on the interaction, which indicates the importance of the Box1 region in the protein interaction.

【基金】 国家自然科学基金资助项目(39670407)
  • 【文献出处】 复旦学报(自然科学版) ,Journal of Fudan University , 编辑部邮箱 ,2003年01期
  • 【分类号】Q754
  • 【被引频次】4
  • 【下载频次】97
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