甲状腺激素受体与Drosha蛋白互作鉴定分析  

Interaction Analysis of Thyroid Hormone Receptor with Drosha Protein

作　　者：尚攀[1] 付元帅[1] 施志仪[1] 喻杰[1] 刘素萍[1] 

机构地区：[1]上海海洋大学农业部淡水水产种质资源重点实验室,上海201306

出　　处：《生物技术通报》2017年第8期206-212,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(41676138;41506159);上海市自然科学基金项目(15ZR1420600);上海市教委科研创新项目(15ZZ083)

摘　　要：为了证明甲状腺激素受体(THR)与Drosha蛋白之间的相互作用,以HEK293细胞为材料,利用免疫共沉淀的方法获得蛋白免疫复合物,再经Western blot检测和质谱分析。结果表明,用THR抗体Western blot检测出有THR蛋白的存在,反之Drosha抗体Western blot检测有Drosha蛋白的存在;同时,THR抗体免疫沉淀后的质谱鉴定存在Drosha蛋白,Drosha抗体免疫沉淀后的质谱鉴定也存在THR蛋白,由此说明甲状腺激素受体与Drosha蛋白之间确实存在相互作用。并且,初步筛选出与两者共同作用的54个蛋白,经Go聚类分析揭示鉴定蛋白主要是细胞膜和细胞核中成分,参与细胞迁移、细胞凋亡、蛋白代谢、免疫应答、信号通路调节及转录调控等生物途径,具有核酸结合、细胞骨架结合等分子功能。This study isto demonstrate the interaction between the thyroid hormone receptor（THR）and the Drosha protein. SelectingHEK293 cells as materials,the protein immune complexes were acquired by co-immunoprecipitation method,and then detected by Westernblot and analyzed by mass spectrometry. The results showed that there was THR protein detected by THR antibody Western blot,andDrosha protein was detected by Drosha antibody Western blot. Concurrently,the Drosha protein was identified by mass spectrometry afterimmunoprecipitation with THR antibody,and the mass spectrometry after Drosha antibody immunoprecipitation confirmed that therewas alsoa THR protein,suggesting that there was interaction between the THR and the Drosha protein. Moreover,the Go cluster analysis for theprimarily screened 54 proteins interacting with both THR and Drosha protein revealed that the identified proteins were mainly composed of cellmembrane and nucleus,and they were involved in varied biological pathways such as cell migration,cell apoptosis,protein metabolism,immune responses,signal pathways and transcription regulations,etc.,also had molecular functions of binding with nucleic acid andcytoskeleton.

关 键 词：甲状腺激素受体 Drosha蛋白 免疫共沉淀 质谱分析 HEK293细胞 

分 类 号：Q51[生物学—生物化学] Q572