Analysis of the Virion Proteins of Human Polyomavirus, BK Virus, by Proteomics-based Approaches

• Main Authors: Hsiang-Ying Chen, 陳香吟
• Other Authors: none
• Format: Others
• Language: en_US
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/43334897343440709164

碩士 === 國立中正大學 === 分子生物研究所 === 93 === BK virus (BKV), a human polyomavirus, infects most of the human population, but clinical disease relevant infections are usually limited to immunocompromised or immunosuppressed individuals. After primary infection, BKV remains latent in the kidney and the urinary tract. Its reactivation causes nephropathy in renal transplant recipients. In order to understand the post-translational modifications of BKV virion proteins VP1, VP2, VP3, and histones, BK virions were propagated in the African green monkey kidney cells (Vero) and purified by sucrose gradient centrifugation. The purified BKV was analyzed by two-dimensional (2D) gel electrophoresis to separate the structural proteins into distinct isospecies. BKV VP1 was identified to be composed of 16 species with pI’s between pH 6.1 and 6.7 by mass spectrometry and Western blot. Analysis of the minor structural proteins revealed only one species of VP3 had pI value of 7.0, and VP2 was not found in this study. By Triton-Acetic acid-Urea (TAU) gel electrophoresis separation and mass identification, the patterns of histone H3 and H4 of BKV virion were different from that of host cells (Vero), especially the subspecies of acetylated H4, and histone H2A and H2B were similar between virus and host cells. The results of 2D and TAU gel electrophoresis may provide important information regarding the post-translational modifications of BKV virion proteins crucial for the viral life cycle.