免疫共沉淀联合质谱对肝细胞核因子3β蛋白复合体的分离鉴定

摘要

目的:利用免疫共沉淀联合质谱分析方法,在人肝癌细胞系细胞株HepG2内筛选与肝细胞核因子(HNF)3β相互作用的蛋白质.方法:通过免疫共沉淀,用特异抗体分离出HNF3β蛋白复合体,质谱鉴定其成分,NCBI数据库检索各个蛋白质的功能,分析筛选与HNF3β相互作用的蛋白质.结果:利用免疫共沉淀联合质谱分析筛选到32个与HNF3β相互作用的候选蛋白质,数据库检索发现LMNA与HNF3β具有相同功能注释,参与转录调控.结论:在细胞内HNF3β与多种蛋白质有相互作用,协同参与转录调控,推测HNF3β/LMNA之间相互作用的变化与葡萄糖及脂肪代谢及相关疾病的发生有关.%Objective: To screen proteins interacting with hepatocyte nuclear factor(HNF) 33 by the method of coimmunoprecipitation coupled with mass spectrometry(IP-MS) in HepG2 cells. Methods: The HNF3|3 complex was purified from HepG2 cells via the method of coimmunoprecipitation with anti-HNF3|3 specific antibody. Then the components of the complex were identified by mass spectrometry. To screen proteins interacting with HNF3|3, the function of each protein was obtained from the National Center for Biotechnology Information (NCBI). Results: 32 proteins interacted with HNF33 were obtained from the study via IP/MS. We found that LMNA and HNF3|$ showed the same fuction involved in the process of transcription. Conclusion: HNF3(J coordinate with a variety of proteins in cells during transcription. We speculate that the interaction between HNF3P and LMNA is related to glucose and lipid metabolism and some diseases.