LTQ-FT-MS技术结合生物信息学筛选与HBV Polymerase存在潜在相互作用的宿主蛋白

发布时间：2018-12-16 23:22

【摘要】：乙型肝炎病毒(Hepatitis B virus,HBV)是慢性肝病的一个主要致病因素,易导致肝炎、肝硬化,并最终导致肝癌。乙型肝炎病毒聚合酶(HBV Polymerase,Pol)在HBV复制过程中起着关键作用,并有可能参与肝癌的发生。Pol的这些生理功能很有可能是通过与蛋白质之间的相互作用来行使的。本文通过免疫共沉淀(Co-immunoprecipitation)结合液相色谱串联质谱(LC-MS/MS)技术,筛选、鉴定与Pol存在潜在相互作用的蛋白。用构建好的pcDNA3.1-POL-flag重组质粒转染HeLa细胞,LC-MS/MS技术筛选能与融合flag标签的Pol发生免疫共沉淀的蛋白,共得到45个蛋白,其中11个蛋白被报道能与Pol发生相互作用。Ingenuity Pathway Analysis(IPAR○)软件进一步分析45个蛋白与Pol的功能及相应的细胞网络,发现这些蛋白涉及三个大分子网络,其中4个蛋白可能参与协助Pol在细胞网络中行使其生理功能,对这4个蛋白的进一步研究将有助于为攻克乙型肝炎病提供线索。
[Abstract]:Hepatitis B virus (Hepatitis B virus,HBV) is a major cause of chronic liver disease, which can lead to hepatitis, cirrhosis, and ultimately liver cancer. Hepatitis B virus polymerase (HBV Polymerase,Pol) plays a key role in the process of HBV replication and may be involved in the development of liver cancer. These physiological functions of Pol may be performed by interaction with proteins. In this paper, immunocoprecipitation (Co-immunoprecipitation) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to screen and identify proteins with potential interaction with Pol. The recombinant plasmid of pcDNA3.1-POL-flag was used to transfect HeLa cells. LC-MS/MS technique was used to screen the proteins that could co-precipitate with the fusion flag tagged Pol. A total of 45 proteins were obtained. Eleven of these proteins were reported to interact with Pol. Ingenuity Pathway Analysis (IPAR0) software to further analyze the functions of 45 proteins and Pol and their corresponding cellular networks, and found that these proteins involve three large molecular networks. Four of these proteins may be involved in assisting Pol to perform its physiological functions in the cellular network, and further study of these four proteins will provide clues for the prevention of hepatitis B disease.
【作者单位】：暨南大学生命科学技术学院;
【基金】：国家科技重大专项课题(2012ZX10002006-003)
【分类号】：R373.21

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