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NS4B蛋白结合的宿主蛋白，制备了组成表达HA标记的NS4B蛋白的人肝癌细胞，并利用光活性氨基酸类似物进行体内交叉。在细胞裂解液中检测到一个约60 KDa的NS4B蛋白与其结合蛋白(S)的交联复合体，并用HA琼脂糖凝胶纯化。然而，该络合物的数量不足以通过质谱学进行鉴定。因此，需要放大提纯过程和/或提高交联率。

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.