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核蛋白MARVELD1与核质转运受体蛋白Importin β1相互作用的鉴定

Identification of Interaction between MARVELD1 and Importin β1
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摘要 核蛋白MARVELD1(MARVEL domain containing1)是一个在多种肿瘤中表达下调的肿瘤相关基因.为寻找其相互作用分子,构建pGEX-4T-2/MARVELD1重组体并在大肠杆菌中成功表达GST-MARVELD1融合蛋白.采用GSTpull-down结合LC-MS/MS(液相色谱-串联质谱)的方法对MARVELD1蛋白的相互作用分子进行筛选,发现了16个与MARVELD1相互结合的蛋白.进一步的免疫共沉淀实验证实,MARVELD1与核质转运受体蛋白importin β1能够相互作用,说明MARVELD1可能参与了importin β1的部分生物学作用,或是它通过与importin β1结合而进入细胞核.研究结果为进一步分析MARVELD1和importin β1的生物学功能奠定了基础. MARVELD1 （MARVEL domain containing 1 ） is a tumor-related gene down-regulated in multiple tumors. To study the molecules interacted with MARVELD1, recombinant pGEX-4T-2/MARVELD1 was constructed and GST-MARVELD1 fusion protein was expressed in E. coll. Sixteen proteins interacted with MARVELD1 were found by GST pull-down followed liquid chromatographytandem mass spectrometry （LC- MS/MS） analysis. The interaction between MARVELD1 and importin β1 was identified by coimmunoprecipitation, which indicates that MARVELD1 may partly participate in the biological function of importin β1 or MARVELD1 was imported into nucleus through binding to importin β1. The results provide important implication for further studies of MARVELD1 and importin β1 function.
作者 王山 肖晟 赫杰 韩放 李钰
机构地区 哈尔滨工业大学生命科学与工程系 美国哈佛大学医学院病理系
出处 《中国生物化学与分子生物学报》 CAS CSCD 北大核心 2009年第8期735-739,共5页 Chinese Journal of Biochemistry and Molecular Biology
基金 国家自然科学基金(No.30170516 No.30871271)资助~~
关键词 MARVELD1 核质转运受体蛋白 GST pull-down 免疫共沉淀 MARVELD1 importin β1 GST pull-down co-immunoprecipitation
分类号 Q71 [生物学—分子生物学] Q503 [生物学—生物化学]
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参考文献20

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中国生物化学与分子生物学报
2009年 第8期

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