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

• Project/Area Number: 21590085
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: National Institute of Infectious Diseases
• Principal Investigator: SAITO Kyoko 国立感染症研究所, 細胞化学部, 主任研究官 (70235034)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2009: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
• Keywords: 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

Research Abstract

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.