FlightlessⅠ与核质转运蛋白Importin β及核孔蛋白Nup88的相互作用  

Interaction of Flightless I with Nup88 and Importin β

作　　者：廖声友 王翠华[1] 汤冬娥[1] 魏金梅[1] 贺玉娇 熊海庭 徐凤梅[1] 高学娟[1] 刘小会[1] 刘朗夏[1] 

机构地区：[1]广东省高校功能蛋白质研究重点实验室暨南大学生命与健康工程研究院,广东广州510632

出　　处：《生物工程学报》2015年第8期1247-1254,共8页Chinese Journal of Biotechnology

摘　　要：Fightless I(FLII)是一个在肿瘤中高表达的蛋白,前期研究提示FLII可能参与核质转运过程,为鉴定FLII是否与核膜结合蛋白相互作用,构建GST-FLII、GST-LRR融合蛋白重组质粒并转化至大肠杆菌Rosetta进行诱导表达,使用GST琼脂糖珠进行融合蛋白的纯化。通过SDS-PAGE对纯化蛋白进行验证之后,利用GST-pull down及免疫共沉淀技术证明了FLII与Importinβ、Nup88蛋白的相互作用,并鉴定FLII上的LRR结构域为相互作用区域。研究结果提示FLII可能参与了Importinβ的部分生物学作用,为进一步分析FLII与Importinβ、Nup88的生物学功能奠定了基础。High expression of Fightless I （FLIl） is associated to multiple tumors. Based on our previous study that FLII might be involved in the nuclear export, we assessed the possible interaction of FLII with the nuclear envelop associating proteins lmportin Ⅰ and Nup88. We first constructed GST-FLII, GST-LRR recombinant plasmids and transformed them intothe Rosetta strain to produce GST-FLII, GST-LRR fusion protein. After purification of these proteins, GST-pull down, as well as co-immunoprecipitation, were used to test the interaction of FLII with Importin β and Nup88. FLII interacted with Importin β and Nup88, and FLII LRR domain is responsible for these interactions. Thus, FLII may play a role in nuclear export through interaction with Importin β and Nup88.

关 键 词：分子生物学 RNA转运 GST-pull down 免疫共沉淀 FLIGHTLESS Ⅰ IMPORTIN β NUP88 

分 类 号：Q78[生物学—分子生物学]