Screening for host factors involved in membrane rearrangements induced by hepatitis C virus NS4B protein

丙型肝炎病毒NS4B蛋白诱导膜重排相关宿主因子的筛选

• 批准号: 21590085
• 负责人: SAITO Kyoko
• 金额: $ 2.16万
• 依托单位: National Institute of Infectious Diseases
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21590085/
• 关键词: ウイルス; 感染症; 生体膜; 宿主因子; ウィルス; C型肝炎ウィルス; NS4B蛋白質

To identify host proteins that bind to hepatitis C virus(HCV) NS4B protein, human hepatoma cells constitutively expressing HA-tagged NS4B protein were prepared and were crosslnked in vivo by using photo-reactive amino acid analogs. A crosslinked complex(ca. 60KDa) of NS4B protein and its binding protein(s) was detected in the cell lysate and purified by using HA agarose. However, the amount of the complex was not enough to be identified by mass spectrometry. Thus, scale up of the purification processes and/or improvement of crosslinking efficiency are needed.

为了鉴定与丙型肝炎病毒（HCV）NS 4 B蛋白结合的宿主蛋白，制备组成型表达HA标记的NS 4 B蛋白的人肝癌细胞，并通过使用光反应性氨基酸类似物在体内交联。交联复合物（CA.在细胞裂解物中检测到NS 4 B蛋白及其结合蛋白，并通过使用HA琼脂糖进行纯化。然而，复合物的量不足以通过质谱法鉴定。因此，需要扩大纯化方法和/或提高交联效率。