Wu S X, Zhu X P, Letchworth G J
Department of Animal Health and Biomedical Sciences, University of Wisconsin-Madison 53706, USA.
J Virol. 1998 Apr;72(4):3029-36. doi: 10.1128/JVI.72.4.3029-3036.1998.
Nine glycoproteins (gB, gC, gD, gE, gG, gH, gI, gK, and gL) have been identified in bovine herpesvirus 1 (BHV-1). gM has been identified in many other alpha-, beta-, and gammaherpesviruses, in which it appears to play a role in membrane penetration and cell-to-cell fusion. We sought to express BHV-1 open reading frame U(L)10, which encodes gM, and specifically identify the glycoprotein. We corrected a frameshift error in the published sequence and used the corrected sequence to design coterminal peptides from the C terminus. These were expressed as glutathione S-transferase fusion proteins in Escherichia coli. The fusion protein containing the 63 C-terminal amino acids from the corrected gM sequence engendered antibodies that immunoprecipitated a 30-kDa protein from in vitro translation reactions programmed with the U(L)10 gene. Proteins immunoprecipitated by this antibody from virus-infected cells ran at 36 and 43 kDa in reducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and 43 and 48 kDa in nonreducing SDS-PAGE. Only the larger of the pair was present in virions. A 7-kDa protein was released from gM by reducing agents. The 7-kDa protein was not recognized in Western blots probed with the anti-gM antibody but reacted specifically with antibodies prepared against BHV-1 U(L)49.5, previously reported to be a 9-kDa protein associated with an unidentified 39-kDa protein (X. Liang, B. Chow, C. Raggo, and L. A. Babiuk, J. Virol. 70:1448-1454, 1996). This is the first report of a small protein covalently bound to any herpesvirus gM. Similar patterns of hydrophobic domains and cysteines in all known gM and U(L)49.5 homologs suggest that these two proteins may be linked by disulfide bonds in all herpesviruses.
在牛疱疹病毒1型（BHV - 1）中已鉴定出九种糖蛋白（gB、gC、gD、gE、gG、gH、gI、gK和gL）。gM已在许多其他α、β和γ疱疹病毒中鉴定出来，它似乎在膜穿透和细胞间融合中起作用。我们试图表达编码gM的BHV - 1开放阅读框U(L)10，并特异性鉴定该糖蛋白。我们校正了已发表序列中的移码错误，并使用校正后的序列从C末端设计共末端肽。这些肽在大肠杆菌中作为谷胱甘肽S - 转移酶融合蛋白表达。包含来自校正后的gM序列的63个C末端氨基酸的融合蛋白产生了抗体，该抗体从用U(L)10基因编程的体外翻译反应中免疫沉淀出一种30 kDa的蛋白质。在还原十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）中，由该抗体从病毒感染细胞中免疫沉淀的蛋白质迁移率为36 kDa和43 kDa，在非还原SDS - PAGE中为43 kDa和48 kDa。这对蛋白中只有较大的蛋白存在于病毒粒子中。还原剂从gM中释放出一种7 kDa的蛋白质。在用抗gM抗体探测的蛋白质印迹中未识别出7 kDa的蛋白质，但它与针对BHV - 1 U(L)49.5制备的抗体特异性反应，先前报道U(L)49.5是一种与未鉴定的39 kDa蛋白质相关的9 kDa蛋白质（X. Liang、B. Chow、C. Raggo和L. A. Babiuk，《病毒学杂志》70:1448 - 1454，1996）。这是关于一种与任何疱疹病毒gM共价结合的小蛋白质的首次报道。所有已知的gM和U(L)49.5同源物中相似的疏水域和半胱氨酸模式表明，这两种蛋白质可能在所有疱疹病毒中通过二硫键相连。
