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

为了鉴定与丙型肝炎病毒（HCV） NS4B蛋白结合的宿主蛋白，制备了组成型表达ha标记NS4B蛋白的人肝癌细胞，并利用光反应性氨基酸类似物在体内交联。交联的络合物(ca。在细胞裂解液中检测到60KDa)的NS4B蛋白及其结合蛋白(s)，并用HA琼脂糖纯化。然而，该络合物的数量不足以通过质谱法进行鉴定。因此，需要扩大净化过程的规模和/或提高交联效率。

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.