Johannsen Eric, Luftig Micah, Chase Michael R, Weicksel Steve, Cahir-McFarland Ellen, Illanes Diego, Sarracino David, Kieff Elliott
Program in Virology, Department of Microbiology, Harvard Medical School and Brigham and Women's Hospital, Boston, MA 02115, USA.
Proc Natl Acad Sci U S A. 2004 Nov 16;101(46):16286-91. doi: 10.1073/pnas.0407320101. Epub 2004 Nov 8.
Mature Epstein-Barr virus (EBV) was purified from the culture medium of infected lymphocytes made functionally conditional for Zta activation of lytic replication by an in-frame fusion with a mutant estrogen receptor. Proteins in purified virus preparations were separated by gradient gel electrophoresis and trypsin-digested; peptides were then analyzed by tandem hydrophobic chromatography, tandem MS sequencing, and MS scans. Potential peptides were matched with EBV and human gene ORFs. Mature EBV was mostly composed of homologues of proteins previously found in a herpes virion. However, EBV homologues to herpes simplex virus capsid-associated or tegument components UL7 (BBRF2), UL14 (BGLF3), and EBV BFRF1 were not significantly detected. Instead, probable tegument components included the EBV and gamma-herpesvirus-encoded BLRF2, BRRF2, BDLF2 and BKRF4 proteins. Actin was also a major tegument protein, and cofilin, tubulin, heat shock protein 90, and heat shock protein 70 were substantial components. EBV envelope glycoprotein gp350 was highly abundant, followed by glycoprotein gH, intact and furin-cleaved gB, gM, gp42, gL, gp78, gp150, and gN. BILF1 (gp64) and proteins associated with latent EBV infection were not detected in virions.
成熟的爱泼斯坦-巴尔病毒（EBV）是从感染淋巴细胞的培养基中纯化得到的。这些淋巴细胞通过与突变雌激素受体的读码框融合，在功能上对Zta激活裂解复制具有条件性。纯化病毒制剂中的蛋白质通过梯度凝胶电泳分离并经胰蛋白酶消化；然后通过串联疏水色谱、串联质谱测序和质谱扫描分析肽段。将潜在肽段与EBV和人类基因开放阅读框进行匹配。成熟的EBV主要由先前在疱疹病毒体中发现的蛋白质同源物组成。然而，未显著检测到EBV与单纯疱疹病毒衣壳相关或被膜成分UL7（BBRF2）、UL14（BGLF3）以及EBV BFRF1的同源物。相反，可能的被膜成分包括EBV和γ-疱疹病毒编码的BLRF2、BRRF2、BDLF2和BKRF4蛋白。肌动蛋白也是一种主要的被膜蛋白，而肌动蛋白结合蛋白、微管蛋白、热休克蛋白90和热休克蛋白70是其主要成分。EBV包膜糖蛋白gp350含量极高，其次是糖蛋白gH、完整的和经弗林蛋白酶切割的gB、gM、gp42、gL、gp78、gp150和gN。在病毒体中未检测到BILF1（gp64）以及与EBV潜伏感染相关的蛋白质。
