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前S1蛋白(PreS1)在乙型肝炎病毒与宿主的相互作用中起至关重要的作用.为筛选乙型肝炎病毒PreS1结合蛋白,进一步探讨其在病毒感染过程中的作用,原核表达、纯化了PreS1-谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)融合蛋白,利用此蛋白与HepG2细胞裂解液进行Pull-down实验,其产物进行双向凝胶电泳分离.结果发现2个PreS1特异结合蛋白,经质谱鉴定为分子伴侣蛋白——葡萄糖调节蛋白78(GRP78)和葡萄糖调节蛋白75(GRP75).通过免疫共沉淀和Western印迹分析证实,PreS1与GRP75之间存在相互作用.实验结果表明,GRP75为新发现乙型肝炎病毒PreS1特异结合蛋白,其与PreS1结合后的生理功能以及在HBV感染过程中的作用值得深入研究.
The pre-S1 protein (PreS1) plays a crucial role in the interaction between Hepatitis B virus and the host.In order to screen the PreS1-binding protein of Hepatitis B virus and further explore the role of PreS1 in virus infection, prokaryotic expression and purification PreS1-glutathione-S-transferase (GST) fusion protein, using this protein and HepG2 cell lysate Pull-down experiments, the product was separated by two-dimensional gel electrophoresis and found that 2 The PreS1-specific binding protein was identified by mass spectrometry as a chaperone of glucose-regulated protein 78 (GRP78) and glucose regulated protein 75 (GRP75) .Immunoprecipitation and Western blot analysis confirmed the presence of an interaction between PreS1 and GRP75. The results showed that GRP75 is a newly discovered PreS1 specific binding protein of hepatitis B virus. Its physiological function after binding to PreS1 and its role in HBV infection deserved further study.